Clinical Biochemistry

Question 1

STP Programme structure

Answer 1

3 year programme that combines full time work with a part time masters.

Intense but rewarding programme.

Training our scientists to get the best of patients and for patient safety.

Will have to travel to the University > out of Ashton, Kings College, Newcastle, Queen Mary, Liverpool, Manchester x2.

We expect out trainees to attend our workplace and also attend the university! This is line with requirements of employer and the uni. NOT distanced learning!

All fees covered.

Application > apply through Oriel.
Passionate about clinical biochem, want to train in and work in, all trainees need resourcefulness, resilience to get through the intense programme.
Demonstrate resilience by reading all the guidance available.
e.g. time for tea youtube channel, lecture at uni, stp perspectives, website.
Consider the person specification and job description > LOOK ON WEBSITE > match my skills and knowledge to these!
Application is your time to shine > think about what you do, your experience / skills that you have and match it to the peron spec + job description. CLINICAL SCIENTIST SHADOWING DAY + PLACEMENT!
How does your current experience compliment what you’re going to learn: understanding what the NHS values are, the focus on patient care, teamwork, confidentiality, respect and how your own experiences so far reflect those qualities! What has led you to pick this career? Teamwork example?

More than 70% of patient healthcare decisions depend on the results of tests carried out in a pathology laboratory. A crucial component in the delivery of high quality patient care, pathology demands accuracy, efficient turnaround and value for money.

EXAMPLES FROM FLEET / JOBS / WORK EXPERIENCE / UNI AND RELATE BACK TO PATIENT FOCUS.

Picking your specialism: Clinical Biochem > I like a challenge and learning new things. Not very good at doing the same thing each day, I thrive off variety and being busy (what I liked about Fleet) which clinical science provides! Varied + new challenges. Understand the day to day role. Time for tea youtube.
I don’t like to be bored and clinical science would ensure I’m not as I would be making a contribution to patient care as well as the opportunity to contribute to research and development also.
Role of the healthcare scientist is becoming more prominent / important > ageing population, unhealthier, obesity rise etc

HOW TO STAND OUT:
Originally, wanted to go into research, however my placement year opened my eyes into the fulfilment that lab-based work can bring.

Have a specific / unique scientific passion that you can showcase

e.g. Year in industry placement at Fleet > placement opened up my eyes into the reality of working in the scientific industry, it is a high pressure, fast paced environment! I learnt that I enjoy that sort of environment as I love keeping busy and a challenge.
It puts the theory I learnt at Uni into practice, puts it into perspective and made me recognise the value of what we learn.

Pepsin digest of HIV and/or Troponin Ab > crucial for healthcare assays / blood tests to work properly to enable accurate diagnosis and treatment of patients!
DNaseB reagent validation testing on Optolite machine > eventually will be used in a hospital lab for patient samples.

Gyros machine project > unique experience using high tech machine > sample prep, authorising and reviewing results, programming and troubleshooting machine > developing a series of ‘fit for purpose’ assays for the quantification of biomarker analyte in human serum. Again, critical that the immunoassay is optimised to accurately identify the cancer biomarker in patients for correct care. Experience in handling human / mouse serum and health and safety implications.

Something relevant to healthcare to demonstrate your passion > e.g. learning about quality at fleet and how it crucially important as eventually the conjugate / assay will end up in a healthcare setting impacting the life of a patient! E.g. antibody, AP, troponin conjugates for use in clinical settings.
Impact on patients > there is a patient at the end of the sample you are testing! The patient is at the centre of everything you do.
At the end of the production line there is a patient and the accuracy of this conjugate impacts the accuracy of their clinical outcomes.

Should be readable and well structured > applicant has high level of curiosity and they embed the NHS values in the way they live their life.
Important to have emotional awareness, it’s a big responsibility, must be a confident leader, works well in a team, communicate ideas effectively, work with challenging patients or colleagues. It is a challenge, it must be something you are wanting to commit to!
Look at the competencies online!

What makes an ideal STP student? (refer to in application / interview, give examples and refer back to NHS patients):
- Good time management
-Self-motivation
- Initiative
- Good resilience
- Competence, curiosity, confidence
-Passion and commitment to applying science to IMPROVE patient care!

Research current issues in healthcare > e.g. antimicrobial resistance in point of care testing.

Situational judgement test:
Practice questions on the pearson view website are the most helpful. Give you feedback on your answers! Familiarise yourself with the NHS values as the test is based around these. Put yourself in a healthcare setting and think what you would do. Be aware of timing!
Pearson view > CAN’T use android or IOS devices > do on computer.

Look at advice on website / STP perspectives.

Interview is online > questions relating to science, leadership, values and behaviours.

Clinical Biochem job description:
The typical work activities that you might undertake include:

planning and organising work in clinical biochemistry laboratories (fleet)
carrying out complex analyses on specimens of body fluids and tissues (Gyros)
assuring the quality of clinical biochemistry investigations (quality -fleet)
auditing the diagnostic and clinical use and performance of investigations (audit at fleet)
developing new and existing tests, often automated and computer assisted but sometimes requiring considerable manual expertise (Gyros)
liaising with clinical and other healthcare staff, often in a multidisciplinary team setting (teamwork at fleet)
some patient contact (contacting customers for shipping at fleet)
writing reports (placement report / uni)
submitting funding bids and conducting research with clinician

Question 2

Clinical Biochemistry

Answer 2

Pathology (the study of disease) includes a number of specialisms, including clinical biochemistry, in which you could help diagnose and manage disease.

In clinical biochemistry, you’ll help to diagnose and manage disease through the analysis of blood, urine and other body fluids.
You’ll do this by producing and validating the results of chemical and biochemical analyses.

1. Apply Principles of Clinical Biochemistry
2. Perform Relevant Techniques
3. Evaluate, Interpret, and Communicate Data
4. Clinical Liaison and Direct Care

Question 3

Day to day activities

Answer 3

Variety and hands on!

Writing competencies - search up

Running assays yourself - sample prep, reviewing results, authorising them. Example at Fleet: Gyros FRD11 (automated machine and prepped assay myself).

Uni work - study days.

Training from other biomedical scientists in department.

Tests you run: lots are automated on big analysers (e.g. sodium + potassium, creatinine, hormones, haematology example), spectrophotometry with immunoassay / enzymology. Sodium and potassium is done with Ion Selective Electrodes.

Whereas, in special chem department it is a lot more manual and hands on: faecal immunochemical tests i.e. little screening samples to look for blood in faeces to determine whether a person may have colorectal cancer. ?
A lot more samples were sent during COVID because they would rather screen first to see if you meet the threshold before referring someone to colonoscopy as that service was very busy at that time!

See how other departments operate on rotations.

Deal with a lot of different organ and disease systems, get a lot of samples coming in from A&E to rule out certain things like heart attacks, glucose monitoring, Hba1c. Lots of variety, interesting!

Mainly lab based with not much patient interaction day to day. May get the chance to work with the point of care team at some point. Occasionally may need to go on a ward round.

Question 4

Useful things to mention in application + advice
PRINT THIS ONE

Answer 4

Remember, the STP offers a unique blend of practical experience, academic learning, and career development, making it an exciting pathway for aspiring scientists like you!

Reading and using SOPs and MBRs on an almost daily basis at Fleet > critical to understand how important quality control is and gain background information into how a controlled environment operates.

First heard about the STP during my year long lab-based placement in a specialist bioconjugation and immunoassay laboratory and further research into the programme made me realise it is exactly the route I want to go down.
Shadowing the scientist’s day-to-day roles and being given my own projects to work on taught me important analytical and research skills needed to be a successful biochemist. Having this experience within the department was very eye-opening and invaluable > motivating me to pursue this specialism.

Want to transfer my scientific passion to a healthcare setting > love the structure of the lab and variety > provide a service / make a difference.

Solidified that a lab based career is what I want to pursue!

Replay enjoyed the split between the practical + theory side of placement and I know the STP programme has a similar format of work based learning and academic programme.

Passion for science + patient care with a goal of improving lives and wellbeing.
Ultimately, want to work in a scientific environment where I am providing a care / service to fulfil a purpose and make a difference!
Would love to work in a clinical based role > directly helping people.
Would love to work in a more lab based role (immunology / biochem), although not opposed to some patient interaction.

Biochem is very broad and varied - a chance to learn a lot of new things + skills which I love!
Also would love to learn how to use the equipment you use in the lab!

Transferable skills - communication, professionalism, teamworking, leadership.
Part time work - Waitrose (working with lots of different types of people, adaptable to different situations), farm shop, yoghurt kiosk, tennis team at Uni.

Relate everything you say to the role / delivering the best service to NHS patients! e.g. this has given me the skills for x, y, z etc… or I did a lab project where I could see how this could help patients in the future for example…
Don’t waste words.
Talk about your experience that is valuable - emphasise it!
Make it as relevant as possible.

Maybe reach out to someone on linked in to gain clinical observation experience.

I can use my knowledge to improve patients’ lives.

Question 5

Interview Questions - Give examples, use STAR technique!

Answer 5

Q. What is clinical biochemistry?
Pathology (the study of disease) includes a number of specialisms, including clinical biochemistry, in which you could help diagnose and manage disease.

In clinical biochemistry, you’ll help to diagnose and manage disease through the analysis of blood, urine and other body fluids.
You’ll do this by producing and validating the results of chemical and biochemical analyses.

1. Apply Principles of Clinical Biochemistry
2. Perform Relevant Techniques
3. Evaluate, Interpret, and Communicate Data
4. Clinical Liaison and Direct Care

Q. Day to day activities of a clinical biochemist?
Variety and hands on!

Writing competencies - search up

Running assays yourself - sample prep, reviewing results, authorising them. Example at Fleet: Gyros FRD11 (automated machine and prepped assay myself).

Uni work - study days.

Training from other biomedical scientists in department.

Tests you run: lots are automated on big analysers (e.g. sodium + potassium, creatinine, hormones, haematology example), spectrophotometry with immunoassay / enzymology. Sodium and potassium is done with Ion Selective Electrodes.

Whereas, in special chem department it is a lot more manual and hands on: faecal immunochemical tests i.e. little screening samples to look for blood in faeces to determine whether a person may have colorectal cancer. ?
A lot more samples were sent during COVID because they would rather screen first to see if you meet the threshold before referring someone to colonoscopy as that service was very busy at that time!

See how other departments operate on rotations.

Deal with a lot of different organ and disease systems, get a lot of samples coming in from A&E to rule out certain things like heart attacks, glucose monitoring, Hba1c. Lots of variety, interesting!

Mainly lab based with not much patient interaction day to day. May get the chance to work with the point of care team at some point. Occasionally may need to go on a ward round.

Q. Can you name 2 tests that we do in biochem?
Liver function tests (group of blood tests that provide valuable information about how your liver is functioning). e.g. Alkaline phosphatase, Bilirubin, Albumin. Abnormal LFT levels prompt further investigation.
Cardiac markers (markers help diagnose and monitor heart-related conditions). e.g. troponin: the most specific and sensitive marker for myocardial damage and creatine kinase: less specific than troponin.
HbA1c test for effective diabetes management: measures your average blood sugar level over the past 2-3 months. It reflects how well your blood sugar has been controlled during that time. Target for diabetes management: Keep HbA1c levels below 7%.

Q. What might alter reference ranges?
- Age, sex, different stages of pregnancy, specimen type.

• Age: As we age, our bodies undergo various physiological changes. These changes can impact the levels of different substances in our blood e.g. partial pressure of oxygen, glucose.
• Sex: Biological differences between males and females lead to variations in reference ranges. e.g. testosterone levels, Haemoglobin concentration, packed cell volume.
• Specimen type (e.g., blood, urine, spinal fluid) and circumstances (fasting, exercise) can also influence reference ranges.
• Pregnancy: unique physiological changes occur. e.g. lower Haematocrit and Haemoglobin Levels and lower serum Creatinine values.

Q. Pick an analyte and how would you measure it?
- Glucose, HbA1c test (crucial for assessing glycaemic control + managing diabetes).
Electrolytes (Na+, K+) on ion selective electrodes (crucial for assessing renal function, endocrine function, and acid-base balance).

Q. General methods used in clinical biochem?
Automated Analysis
ISE
Immunoassay
Mass spectrophotometry > identifies MW of proteins.
Electrophoresis
Chromatography > Used size exclusion and affinity chromatography on the AKTA columns at Fleet for protein purification.
DNA technology > polymerase chain reaction, gene sequencing.

Q. Introduce yourself and tell me why you want to work for the NHS?
- I am hard working, standards driven person. I enjoy working in a challenging environment where everyone in the team is striving to achieve the same goal of achieving outstanding patient care.
- I have built up extensive experience during part time jobs and my year in industry. The qualities and skills I have developed are closely aligned to this role.
- I want to work for the NHS as the core values are a match for my own. WORKING TOGETHER FOR PATIENTS, DEMONSTRATING COMPASSION, RESPECT AND DIGNITY, A COMMITMENT TO QUALITY CARE, HELPING TO IMPROVE PEOPLE’S LIVES and EVERYONE COUNTS are all values I strongly believe in.
- I am a positive, enthusiastic person, I work well under pressure and I take my professional development seriously as demonstrated throughout my placement year. (PDR).
- If you hire me for this role, you will be employing someone who can contribute positively to the NHS team whilst delivering first-class service to your patients!

Q. What are the top 5 competencies that are critical in this Band 5 position?
After having studied the job description, the competencies required for this position, in my opinion, are:
Technical Competence > without this, you cannot carry out your work to the necessary standards or achieve optimum patient care.

Outstanding Communication Skills > you must have these but also be aware that communication is a two-way process for it to be effective. It is about listening as well as speaking.

Demonstrating Care, Compassion + Patience > you must be understanding of people’s situations when giving advice, information or treatment.

Understanding of Health, Safety and Security > you must be aware of your surroundings, report incidences that compromise safety (e.g. Fleet and lifting heavy weights to calibrate the balances) or breach the operational guidelines.

Equality and Diversity > another very important competency. Everyone who works in the NHS must act in a manner that supports equality and diversity. This means treating people fairly, appropriately, with respect and in line with their own specific needs.

Q. Describe a situation where you used ‘best practice’ in your work?
Fleet example > Everyday in the lab it is important to operate using best lab practice at all times to maintain the controlled environment. As many of the bioconjugation and immunoassay products made at Fleet bioprocessing end up in in-vitro diagnostic products and pre-clinical environments, it is critical that best practice in the lab is used at all time to produce high quality products.
E.g. Must abide by QMS basic principles: Controlled Documents, Quality Records and Traceability. General examples include following MBRs and SOPs, trained staff, using qualified equipment, wearing PPE, monthly lab cleaning, Bioburden monitoring.

Q. Tell me about a time where you challenged someone because of substandard care or compassion?
Example > Part time job at Waitrose and sandwich order problem, all the sandwiches that arrived were out of date. Colleague on the welcome desk was very unhelpful to the customer and was being rude to them. I stepped in, alerted the manager and found a solution to the problem, acting with compassion and understanding at all times. Took responsibility for the substandard customer service my colleague was providing.

Q. Tell me about a time where you received feedback from someone based on your performance on your attitude?
Fleet example > During my placement year I had an April appraisal where I received feedback from the company directors on the progress I had made in my first 8 months of my placement. The feedback I received was positive, they were really pleased with my willingness to learn and adapt, my curiosity and hard working, enthusiastic nature. They were so happy with my attitude and performance that I even received a pay rise!

Q. Tell me about a time when you shared your skills, knowledge and expertise with someone less experienced than you?
Examples > Occasions where I have had to train up new employees.

Fleet - it was my responsibility to train up the lab assistant who would be taking over my role. Duties I had to handover included cleaning and maintenance of the lab, buffer making, equipment calibrating, weekly fire alarm testing, lab coat cleaning and admin jobs such as scanning of buffer sheets and bioburden monitoring.

Farm Shop and Frozen Yoghurt Kiosk - As I was the most experienced employee, it was my job to train up the new starters on the basic features of the job such as cleaning, replenishment, stock checks, customer service requirements.

Training new employees requires patience, respect and dignity, compassion and outstanding communication and listening abilities to make them feel at ease and comfortable at all times.

Q. What are the 6 NHS values and what do they mean?
Working Together for Patients: This value emphasises that patients come first in everything the NHS does. It involves collaboration among healthcare professionals to provide the best care possible.

Respect and Dignity: The NHS values every person, whether they are patients, their families, or staff. This means treating everyone with respect, valuing their opinions, and maintaining their dignity.

Commitment to Quality of Care: The NHS strives to provide high-quality care by ensuring safety, effectiveness, and a positive patient experience. This involves continuous improvement and learning from feedback.

Compassion: Compassion is central to the care provided by the NHS. It means responding with kindness and humanity to each person’s pain, distress, anxiety, or need.

Improving Lives: The NHS aims to improve the health and wellbeing of people and their experiences of the healthcare system. This involves striving for better health outcomes and enhancing the quality of life.

Everyone Counts: This value ensures that resources are used for the benefit of the whole community, and that no one is excluded, discriminated against, or left behind.

Q. What is the NHS constitution?
HIGH QUALITY HEALTHCARE AVAILABLE FOR ALL

Places patients at the heart of everything.
Collaboration.
NHS remains accountable to the public and patients it serves.

For patients and those using NHS services, the NHS Constitution emphasizes the following principles:

The NHS provides comprehensive services available to all based on clinical need, not an individual’s ability to pay.

Aspiring to excellence and professionalism, the NHS places patients at the heart of everything it does.

Collaboration across organizational boundaries and partnerships with other entities serve the interests of patients, local communities, and the wider population.

The NHS remains accountable to the public, communities, and the patients it serves.

*These values shape the NHS’s commitment to delivering high-quality healthcare and ensuring equitable access for all.

Q. What are the six C’s of care in the NHS?
Care
Competence
Compassion
Courage
Communication
Commitment

Q. What are the 3 core principles of the NHS?
When the NHS was founded in 1948, it was launched on three core principles – that it meets the need of everyone, that it be free at the point of delivery, and the care given be based on clinical need, not the ability to pay.

Even now, these principles are the guiding values that drive the NHS and the care they provide.

Q. What Qualities Make a Good NHS Employee?
Focus your answer on the qualities that the six core values of the NHS demand:

Integrity and accountability – To ensure patient confidentiality, learn from mistakes and work towards continuous improvement

Flexibility and time management – To meet ever-changing demands and respond effectively in a challenging environment

Kindness, compassion and patience – To provide the highest standards of patient care and to take an empathetic approach to the treatment of family, friends and carers

Teamwork and communication – To work as part of a healthcare network that includes multiple departments and external organisations

Passion and commitment – To uphold the standards of the NHS and go the extra mile to improve the lives of others.
E.g. when my car broke down for a week, I went the extra mile to ensure I still made it to work.

Q. What is safeguarding and and how would you implement it into your daily work in the NHS?
Safeguarding refers to the measures taken to protect the health, well-being, and human rights of individuals, especially those who are vulnerable, such as children, elderly people, and those with disabilities.

Safeguarding is a fundamental aspect of healthcare that involves protecting individuals, particularly vulnerable groups, from harm, abuse, and neglect. In the NHS, safeguarding is crucial to ensure that all patients receive safe and effective care.

To implement safeguarding in my daily work, I would:

Stay Informed and Trained
Recognise Signs of Abuse or Neglect
Report Concerns Promptly
Maintain Confidentiality
Promote a Safe Environment
Collaborate with Colleagues
Empower Patients.

Q. Give an example of when you provided treatment or care that was in line with one of the 6 NHS values?
Compassion > Could give an example of when I volunteered at Millbrook RSPCA and had to spend time observing and checking the vital signs of a litter of sick kittens that had come into the centre. I had to dedicate the entire day to monitoring them, administering medication and food and making another staff member aware if their condition started to deteriorate.

This experience reinforced the importance of compassion in healthcare, as it not only improves patient outcomes but also enhances their overall well-being.

Q. What are your strengths that will enable you to perform to a high standard in this NHS role?
During my placement year and part time jobs, I have developed several strengths that I believe will enable me to perform to a high standard in this NHS role:

Strong Communication Skills: I excel in both verbal and written communication, which is essential for effectively interacting with patients, families, and colleagues. Clear communication ensures that patient care is coordinated and that everyone involved is well-informed.

Empathy and Compassion: I have a deep sense of empathy and compassion, which allows me to connect with patients on a personal level. This helps in providing care that is not only clinically effective but also emotionally supportive, ensuring patients feel valued and understood.
e.g. demonstrating compassion to pet owners at vet practices during emotional situations.

Attention to Detail: I am meticulous in my work, ensuring that all procedures are followed accurately. This attention to detail helps in minimising errors and providing high-quality care.
e.g. Quality records at Fleet such as following MBRs, SOPs carefully and keeping calibration records and lab book write ups up to date. Gyros project required high attention to detail when formulating 132 samples.

Team Collaboration: I thrive in a team environment and believe in the power of collaborative work. I have experience working with multidisciplinary teams, and I understand the importance of each team member’s role in delivering comprehensive patient care.
e.g. working as a team at Waitrose, the farm shop and monthly lab cleaning at Fleet.

Adaptability: The healthcare environment is dynamic and often unpredictable. I am adaptable and can handle changing situations with ease, ensuring that patient care remains uninterrupted and of high quality.
e.g. at Fleet it was common for me to be given short notice on multiple tasks that needed doing if they were urgent, I had to be adaptable to manage my time between both departments as well as still fulfilling my lab maintenance duties.

Commitment to Continuous Learning: I am dedicated to my professional development and regularly seek out opportunities for training and education. This commitment ensures that I stay updated with the latest healthcare practices and can provide the best possible care to patients.
e.g. further reading into NHS current research such as new Epicbeaker IT system, drone delivery for drugs, CRP being used in POCT and higher sensitivity troponin test. Clinical scientist shadowing day!

Problem-Solving Skills: I have strong problem-solving abilities, which enable me to quickly assess situations, identify issues, and implement effective solutions. This is crucial in a healthcare setting where timely decision-making can significantly impact patient outcomes.
e.g. when lab work did not go to plan at Fleet it was vital to adapt quickly to problems and find solutions > plate washer calibration malfunction.

By leveraging these strengths, I am confident that I can contribute positively to the NHS team and provide exceptional care to patients.

Q. What knowledge do you have of NHS systems or processes?
NHS Systems and processes:

1. Overview of NHS Structure
   Primary Care: This includes services provided by general practitioners (GPs), dentists, pharmacists, and optometrists.

Secondary Care: This involves specialist services provided by hospitals, including emergency care etc.

Tertiary Care: Highly specialised care, often for complex conditions, provided in specialised hospitals or units.

2. Key NHS Systems:
   Electronic Health Records (EHR): Digital versions of patients’ paper charts
   Patient Safety Incident Response Framework (PSIRF): Managing and learning from patient safety incidents
   Quality Management Systems: Ensuring continuous improvement in care quality
3. Processes and Protocols:
   Patient Pathways: Understanding the journey a patient takes from initial consultation through to treatment and follow-up, ensuring continuity of care.

Clinical Governance: Knowledge of frameworks that ensure patient safety, quality of care, and continuous improvement, including audits, risk management, and incident reporting.

Data Protection: Familiarity with GDPR and the Data Protection Act, ensuring patient information is handled securely and confidentially.

4. Experience and Application:
   Practical Experience
   Training and Development.

Q. What is the biggest hazard in the NHS?
Risk of infection from handling patient blood samples > e.g. HIV, hepatitis etc. Could accidentally transmit virus by touching needle.

Health and safety precautions > at Fleet had to sign off SOP / documents before handling human samples and was warned about H+S implications.

What measures do you have in place to protect staff from potential infections when handling blood samples?

Certified samples vs uncertified:
Certified = screened for viruses such as hepatitis and HIV before use = safer.
Uncertified = not screened before use = not as safe.

Q. What Are the Current Challenges Facing the NHS?
Funding, staff shortages, rates of pay, working conditions and the allocation of resources are all challenges for the NHS. You must be up to speed on the particulars of each at the time of interview.

Beyond financial and political challenges, there are also many social factors affecting the NHS, such as an ageing population and the impact of poor lifestyle choices, including the rise of diseases like diabetes and obesity.

Q. What would you do if a patient under your care started to get upset or confused?
“If a patient under my care started to get upset or confused, I would first ensure that I remain calm and composed, as this helps to de-escalate the situation. I would use open body language and maintain eye contact to show that I am attentive and concerned.

I would listen actively to the patient, allowing them to express their feelings without interruption. I would acknowledge their emotions by saying something like, ‘I can see that you’re upset, and I’m here to help.’
Providing reassurance is key, so I would explain clearly and simply what is happening and what steps we will take to address their concerns.

I would assess the situation to identify any triggers and evaluate if there are any immediate needs or changes in the patient’s condition. Depending on the situation, I might offer comfort measures such as a glass of water or a quiet space. If necessary, I would involve other healthcare professionals to provide additional support.

Finally, I would monitor the patient to ensure they are feeling better and that their concerns have been addressed. I would also document the incident in the patient’s notes, detailing what happened and the actions taken to ensure continuity of care.”

Q. What is your biggest weakness?
“One of my biggest weaknesses is that I can be overly detail-oriented. While attention to detail is important, I sometimes find myself spending too much time on minor aspects of a project, which can affect my overall efficiency.

I recognized this tendency early in my career when I noticed that I was often the last to finish tasks, even though the quality of my work was high. To address this, I started using time management techniques, such as setting specific time limits for each task and prioritising tasks based on their impact.

I’ve also learned to step back and look at the bigger picture, ensuring that I focus on what truly matters. This approach has helped me become more efficient without compromising the quality of my work. While I still value attention to detail, I now balance it with a broader perspective, which has improved my productivity and effectiveness.”

Q. What is clinical governance and how would you adhere to it within this role?
“Clinical governance is a framework that ensures the NHS delivers high-quality care by focusing on patient safety, clinical effectiveness, and patient experience. It involves continuous improvement and accountability in healthcare delivery.

To adhere to clinical governance in my role, I would prioritise patient safety by following established protocols and reporting any incidents or near misses. I would stay updated with the latest clinical guidelines and best practices to ensure clinical effectiveness. Engaging with patients, listening to their concerns, and involving them in their care decisions would be key to enhancing patient experience.

I would also commit to continuous professional development, supporting my colleagues, and fostering a positive team environment. Participating in clinical audits and quality improvement initiatives would help me identify areas for improvement and implement necessary changes. Lastly, I would handle patient information with the utmost confidentiality and security, adhering to GDPR and NHS policies.

By integrating these practices into my daily work, I would contribute to the overall goal of providing safe, effective, and high-quality care to patients.”

Q. How will you handle the stress and pressure that comes with this role?
“I understand that working in the NHS can be stressful and demanding, but I believe I am well-equipped to handle the pressure. I prioritise effective time management, ensuring that I stay organised and focused on my tasks. I also practice mindfulness and relaxation techniques, such as meditation, to stay calm and centred.

Maintaining a healthy lifestyle is important to me, so I make sure to exercise regularly, eat a balanced diet, and get enough sleep. This helps me build resilience against stress. I also value teamwork and believe in supporting my colleagues. Effective communication and collaboration can significantly reduce pressure.

Taking regular breaks and ensuring I have downtime to relax and unwind outside of work is crucial to avoid burnout.

In my previous roles, I have successfully managed stress and pressure by staying adaptable and maintaining a positive mindset. I focus on the rewarding aspects of the job and celebrate small victories. By implementing these strategies, I am confident in my ability to handle the stress and pressure that comes with this role in the NHS.”

Q. Give me a situation you were in when where you demonstrated a total commitment to the quality of care?
Situation: During my time on work experience at an equine hospital, I encountered a horse who had recently undergone surgery to treat colic and was experiencing significant post-operative pain.

Task: My responsibility was to ensure that the horse received the highest quality of care, which included managing their pain effectively and monitoring their recovery.

Action: I noticed that the horse’s pain medication was not providing sufficient relief. I promptly communicated with the attending vet to discuss alternative pain management options. Additionally, I spent extra time with the horse, providing comfort and reassurance.

Result: As a result of these efforts, the horse’s pain was brought under control, and their recovery process improved significantly.
The owner was extremely pleased with the high level of care that was dedicated to their pet.

OR Situation at Fleet:
Dave and Hypoglycaemic Episode.

One of the directors suffers with type 1 diabetes and was notoriously bad at taking his medication / monitoring his blood glucose levels. There was an occasion when I was the only one in the room with him when he had a Hypoglycaemic Episode. I quickly went to his aid, making he sure he was sitting down and comfortable. Following his advice he had told all staff at Fleet, I grabbed energy gels from his desk and put them near him in case he needed them. Then I gave him space so that he felt comfortable but stayed near enough in case I needed to intervene again by calling 911. Providing a high quality of care was vital here to ensure that Dave’s symptoms did not persist for long and did not worsen which could’ve then lead to serious consequences.

Q. Teamwork and Communication are very important for working in NHS can you give and example of when you use one or both?

I once coordinated a group lab clean while on placement where we divided tasks based on strengths and held regular check-ins to ensure everyone was on track. This improved our efficiency and the quality of our work.

Q. If you overheard a member of the healthcare team talking in an inappropriate manner to a patient what would you do?
Task: My responsibility would be to ensure that all patients are treated with dignity and respect, and to address any behaviour that compromises the quality of care.

Action: I would approach the situation calmly and professionally. I would intervene by politely asking my colleague to step aside for a moment. I would speak to them privately, expressing my concerns about their behaviour and reminding them of our commitment to patient respect and care. I would also offer to assist with the patient if they were feeling overwhelmed.
Afterward, I would check in with the patient to ensure they were okay and apologise for the incident, reassuring them that their concerns were being taken seriously.

Q. How would you deal with a difficult or aggressive patient?
Task: I would first try to de-escalate the situation, ensure the patient’s safety, and provide the necessary care.

Action: I would approach the patient calmly and maintain a non-threatening posture. I would listen actively to their concerns without interrupting, acknowledging their frustration and pain. I would reassure them that their concerns were valid and that I was there to help. To further ease the situation, I would offer to provide updates on their status regularly and ensure they had access to any immediate comfort measures, such as a blanket or water.

By maintaining a calm and empathetic approach, hopefully this would enable me to build trust and cooperation. This would ensure that the overall atmosphere in the emergency department remained stable and safe for all patients and staff.

Q. Tell me everything you know about infection control?

Key Principles of Infection Control
Hand hygiene: Regular and proper hand-washing.
Personal protective equipment (PPE): Use of gloves, masks, and gowns
Safe management of care environment and equipment: Cleaning and disinfecting surfaces and tools
Waste disposal: Proper disposal of medical waste and sharps
Patient placement: Isolating patients with infectious diseases

Sterilisation: Complete elimination of all forms of microbial life, typically used for surgical instruments.
Disinfection: Reducing the number of pathogenic microorganisms to a safe level, used for surfaces and equipment that do not require sterilisation.

Vaccination:
Role: Vaccinations protect healthcare workers and patients from vaccine-preventable diseases.

Importance of Infection Control
Patient Safety
Healthcare Worker Safety
Public Health: Prevents the spread of infections within the community, especially during outbreaks.

Challenges in Infection Control
Compliance
Resource Limitations
Emerging Pathogens: Adapting to new and emerging infectious diseases, such as COVID-19.

Q. What is the professional duty of candour?
The professional duty of candour is a fundamental ethical responsibility for healthcare professionals. It requires them to be open and honest with patients when something goes wrong with their treatment or care. Here are the key aspects:

Transparency: Healthcare professionals must inform the patient (or their advocate, carer, or family) when an error occurs that causes, or has the potential to cause, harm or distress

Apology: They must offer a sincere apology to the affected person

Remedy and Support: Professionals should provide an appropriate remedy or support to rectify the situation if possible

Explanation: A full explanation of what happened, including the short and long-term effects, should be given

Internal Openness: Professionals must also be honest with their colleagues, employers, and relevant organisations, participating in reviews and investigations when necessary

This duty ensures that trust is maintained between healthcare providers and patients, fostering a culture of safety and continuous improvement.

Q. Tell me how you would contribute to the continuous improvement of the NHS trust in this role?
To contribute to the continuous improvement of the NHS trust in this role, I would focus on several key areas:

Patient-Centered Care: I would prioritise patient feedback and experiences to identify areas for improvement. By actively listening to patients and their families, I can help implement changes that enhance the quality of care and patient satisfaction.

Collaboration and Teamwork: Working closely with colleagues across different departments is crucial. I would foster a collaborative environment where ideas and best practices are shared, ensuring that we learn from each other and continuously improve our services.

Professional Development: Staying updated with the latest advancements in healthcare and participating in ongoing training and education is essential. I would commit to continuous learning and encourage my team to do the same, ensuring we are always equipped with the best knowledge and skills.

Quality Improvement Initiatives: Actively participating in and leading quality improvement projects would be a priority. By using data and metrics to identify areas for improvement, we can develop targeted strategies to address these issues and monitor progress over time.

Open Communication: Maintaining open and transparent communication with all stakeholders, including patients, staff, and management, is vital. This ensures that everyone is informed, engaged, and working towards common goals.

By focusing on these areas, I believe I can make a meaningful contribution to the continuous improvement of the NHS trust, ultimately enhancing the quality of care we provide to our patients.

Q. Where do you see yourself in 5 years?
In five years, if I am successful in this NHS role, I envision myself having grown both professionally and personally, contributing significantly to the trust’s goals. Here are some specific aspirations:

Advanced Expertise: This would involve continuous learning and possibly pursuing further qualifications or specialisations.

Leadership Role: I see myself taking on more leadership responsibilities, perhaps in a managerial or supervisory capacity.

Innovation and Research: I would like to be involved in innovative projects or research that contribute to the advancement of healthcare practices. This could include implementing new technologies or participating in clinical trials that bring cutting-edge treatments to our patients.

Personal Growth: On a personal level, I hope to have developed a balanced approach to work and life, maintaining my passion for healthcare while also ensuring my well-being and resilience.

Q. Why is quality so important in the lab?
* Critically important that products work as expected > giving customers what they asked for / producing accurate and reliable results.
* At the end of a product timeline a human beings health and life depends on the product working as expected > e.g. the product could end up in a hospital lab. Quality measures mean checking every step as you go along to ensure no faults in the final product.
* Example at Fleet – T3 Gelatine: Used for thyroid tests > if conjugate isn’t right the test may give the wrong result > patient may be prescribed wrong medication or falsely diagnosed > direct impact on people’s lives.
* NCNs (non-conformance notes) > prevent the process going wrong again.
* Ultimately, the purpose of quality is to keep the customers happy > do it right every time to the best of your ability.

Q. Give an example of working in a high pressure environment?
At Fleet I had to learn to juggle my tasks between both chemistry and assay departments. Good time management and communication with my placement supervisor was essential to balance the high workload and manage my stress in the high pressured lab environment.

E.g. STP > Clinical Biochem will provide samples to haematology and haematology will provide samples to immunology etc.

The links between departments are also key to provide high quality patient care.

Q. What is good scientific practice?

Patients are the number one priority and their safety in all settings where the healthcare science workforce
operate is paramount.
The purpose of Good Scientific Practice (GSP) is to set out the professional standards on which safe and good working practice is founded for all those in the healthcare science workforce.

The standards are essential for all members of the workforce to perform their job role activities; provide safe, effective patient care; and demonstrate their professionalism.

Q. What constitutes a good leader?
Self awareness
Treating people with respect
Good communication and friendly / approachable nature
Actively listening and providing feedback
Value your colleague’s knowledge and skills and address when development is needed.
Celebrating team success.
Being open to new ideas / having a growth mindset
Acting as a role model and leading by example.
Inspiring a positive work ethic / environment.

Q. Clinical Biochem - what is expected of me?
Patients that come into a hospital may have a blood sample taken, and some time later they’ll get their results back, with little knowledge about the processes that have occurred between these points in order to produce these result!

But the steps in between depend upon the work of clinical biochemists and a wide range of laboratory staff, each having a crucial role in the analysis of a sample.

Biochemical analysis has been crucial in the detection and treatment of diseases, including diabetes, inherited disorders/diseases, deficiencies, cancers, and many other illnesses.

In addition to routine work, a clinical biochemist also has a significant role in the development and validation of new tests being introduced in the department.
This ensures that the services available in the laboratory are of the highest standard and will ensure the most accurate clinical outcomes to the patient.

A clinical biochemist may also partake in research in the department.

It is important to know that STP clinical biochemistry specialism is more lab based and therefore has less patient interaction in comparison to other STP specialities!

Q. What does a day look like as a trainee biochemist?
Currently most of the day-to-day work revolves around learning competencies, while also juggling learning the university content.

Some of the competencies I’ve worked towards so far include receiving, labelling, and storing samples, and more recently moving onto analysing the samples myself, using a wide range of techniques such as flow cytometry, mass spectrometry, and many more.

It’s also important to learn about how these analysers work to help with trouble shooting and maintenance, and how to read and report the results of the test.

Example - different types of blood samples:

Different samples require blood to be stored differently depending on the test. This is indicated with the different colour caps.

The purple tube can be used for a full blood count, measuring the amount of red blood cells, white blood cells and platelets that are present in the blood.

The green tube is for serum samples, that allows us to measure analytes like urea and electrolytes.

While the blue tube is used to screen the coagulation of a blood sample (the colours may vary in different hospitals!).

These tests allow us to pick up abnormalities in a patient’s blood sample, and can assist with the diagnosis of a wide range of diseases.

This has a big impact on patient outcomes, making the work of a biochemist vital for the NHS.

Q. What diseases are key to the clinical biochem speciality?
Liver diseases
Kidney disorders
Cardiovascular diseases (heart)
Diabetes mellitus
Cancer

Inherited metabolic disorders e.g. PKU - Phenylketonuria > a rare inherited disorder that affects how the body breaks down protein. People with PKU cannot break down the amino acid phenylalanine, which then builds up in their blood and brain. This can lead to brain damage, seizures and disabilities.

Hypertension (high blood pressure)
Obesity > + related health issues

Accurate diagnosis, treatment and monitoring relies on biochemical tests.

Q. How would you deal with a discrepancy in your lab data?
If I noticed a discrepancy in my data, the first thing I would do is take a step back and assess the situation.
I would review all of the data to determine what caused the discrepancy and if it was an isolated incident or part of a larger pattern.
Once I had identified the source of the discrepancy, I would then work to find a solution. Depending on the nature of the issue, this could involve re-running experiments, revising protocols / MBRs, or further analysing the data.

During my time on placement, I have encountered a few data discrepancies and learnt how to identify and resolve them.

EXAMPLE > Gyros project: Testing of a protein-drug analyte for a biopharmaceutical company. Encountered a few errors when working on this project e.g. one assay run did not generate a standard curve, perhaps due to errors in making up the standards on my apart due to insufficient training and pipette inaccuracy.
To overcome this, I requested more training on correct pipette technique for tiny volumes and ran the assay again, taking extra care this time and thankfully, the second run worked! Important to request extra training when unsure, don’t be scared to ask and always pay attention to detail.

My attention to detail and problem-solving skills make me well-suited for this role.

Q. How would you deal with a difficult patient?
Dealing with difficult patients can be challenging, but it’s an essential part of providing quality healthcare:

Stay calm and professional at all times.

Listen to what the patient is saying > empathise and understand.

Put yourself in their shoes to try and understand their point of view.

Communicate effectively.

Assess the situation > self reflect and don’t let personal bias or stressors come into the equation!

Q. Why do you want to work in lab / for NHS?
* More than 70% patient healthcare decisions depends on the results of tests carried out in a laboratory.
* Crucial component to deliver high quality patient care
* NHS values
* Importance of accuracy, efficiency and attention to detail.
* Help to save lives and make a difference.
* Forefront of research + innovation = patients continually receive the very best healthcare.

Q. What automated analysers do they use in Clinical Biochem and what do they test for?
Cobas 8000 analyser - Tests:

• Ion-selective electrode (ISE)module is used to measure
  the concentration of electrolyte such as Na+ and K+ in the
  sample by measuring the electrical potential difference.
• General chemistry module (c702) is used for measuring a
  wide range of analytes such as glucose, creatinine (kidney) and C
  reactive protein (liver = infection / inflammation) (CRP). Concentration of the analytes is
  measured with the colour intensity of the reaction mixture.
  Creatinine level in the blood or urine shows how well the kidneys are working
• Immunoassay module (e602) measures the concentration
  of analytes such as vitamin B12, ferritin using immunoassay.

Sebia capillary 3 TERA analyser:

*This analyser is used for measuring
the amount of glycated haemoglobin
(HbA1c) in patients’ samples.

• HbA1c can reflect the glucose level in
  patient’s blood over the past 1-2
  months.
• The analyser measures the HbA1c
  concentration by the capillary
  electrophoresis system.
• The measurement of HbA1c aids the
  diagnosis and monitor of diabetes
  mellitus.

Q. What are the QMS basic principles? Quality measure.
1. Controlled Documents - such as SOPs, MBRs. These are compliant with ISO quality standards. Must make sure you use the current versions of these documents.

2. Quality Records - document the output of all procedures e.g. paperwork, lab book write up, equipment calibration records, scanning buffer sheets.
3. Traceability - means you have documented enough detail within our quality records to show exactly how the procedure was followed e.g. recording lab book write ups.

Q. Current research in the NHS?

Point of care testing team (POCT) in the NHS added a new blood test called C-reactive protein (CRP) >
CRP is a protein made by the liver and levels increase when there is inflammation or infection in the body > CRP is one of the best indicators of inflammation in the body!
CRP testing empowered timely decision-making for antibiotic prescriptions. Instead of waiting for lab results, the teams could make real-time decisions > prevented 13 admissions to hospital!

New higher sensitivity troponin test >
High-sensitivity troponin tests can detect lower levels of troponin T in the blood earlier than standard tests. They rule out of a specific type of heart attack called NSTEMI (non-ST-segment elevation myocardial infarction) earlier. Reduces hospital admissions!

Antimicrobial resistance poses a significant threat to the NHS>

AMR occurs when microorganisms (such as bacteria, viruses, fungi, and parasites) become resistant to antimicrobial medicines like antibiotics etc.
The World Health Organization (WHO) has declared AMR as one of the top 10 global public health threats.
Challenges include: longer hospital admissions, increased risk surgeries and higher death rates.
Healthcare strategies to tackle resistance: early prevention and timely diagnosis > relies on the work of the biochemists!

Drone delivery in the NHS >
In 2022, the NHS announced a ground-breaking trial where cancer patients would receive chemotherapy drugs via drones.
Drone delivery takes 30 mins compared to the usual 4 hours. This efficiency benefits patients and minimizes the risk of drug deterioration due to short shelf life.
The success of this trial could pave the way for same-day orders of vital medical treatments. This demonstrates the NHS’s commitment to ensuring timely treatment while also cutting costs and carbon emissions.
Drone technology could help England become a world leader in cancer care!

Epic Beaker IT System all over the NHS >
In 2023, Guy’s and St Thomas’ NHS Foundation Trust and King’s College Hospital NHS Foundation Trust jointly launched the Epic Beaker system.
This was the largest single go-live of Epic’s electronic patient record anywhere in the world.
It is one single, electronic IT system = It provides staff with a complete overview of a person’s care, freeing up more time for patient care.
Benefits: comprehensive data, efficiency and patient empowerment using MyChart app.
The successful implementation of Epic Beaker represents a significant step forward in healthcare technology, benefiting both patients and staff!

Q. What is Point of Care Testing?
Point of Care Testing (POCT) in the NHS refers to conducting diagnostic tests at or near the location of the patient, rather than in a conventional laboratory setting.

The goal of POCT is to collect specimens and obtain accurate results quickly, allowing for immediate clinical decisions!

These tests typically involve blood and urine testing.

The Point of Care Testing (POCT) team brings benefits to patients in allowing timely decision-making, often permitting life-saving interventions to be administered immediately = better care experience for patients!

Q. Skills you gained from placement?
Valuable for my professional development.
Accuracy, efficiency and attention to detail.
Laboratory experience
Diversified experience working between 2 departments > immunoassay + Chem
Experience in automated and manual immunoassay (ELISA) (Gyros)
Troubleshooting / calibrating laboratory equipment
Assay optimisation (FRD11)
Quality management systems (QMS) experience > Audit and experience of quality control measures (NCNs etc)
Technical skills gained in lab techniques > understanding of analytical techniques.
Knowledge of health + safety procedures (meetings, fire alarms, monthly check ups, PPE).
Presentation skills > research ability + competence (fleet presentations and university).

Q. What is an autoimmune response?
The body’s own cells have proteins on their surface, too. But those proteins don’t usually trigger the immune system to fight the cells.

Sometimes the immune system mistakenly thinks that the body’s own cells are foreign cells. It then attacks healthy, harmless cells in the body.
This is known as an autoimmune response.

Examples of autoimmune diseases: Crohn’s disease, Lupus, Type 1 diabetes.

Q. What is an antigen?
The immune system can be activated by a lot of different things that the body doesn’t recognize as its own.

These are called antigens. Examples of antigens include the proteins on the surfaces of bacteria, fungi and viruses.

Q. How does the immune system change in response to allergies?
When a person who is allergic to a substance encounters it, the immune system may react by producing antibodies that “attack” the allergen.
This can cause wheezing, itching, runny nose, watery or itchy eyes, and other symptoms.

The immune system will produce immunoglobulin E, IgE, antibodies for each allergen.
The antibodies will cause cells in the body (mast cells + basophils) to produce histamine.

Once the allergen binds to IgE, specific types of cells (Mast Cells) will release chemicals that trigger the symptoms of the allergic reaction.

Q. What are antibodies?

What are epitopes?
Antibodies are proteins produced by the immune system to help fight off pathogens.

In more scientific terms, antibodies neutralise pathogens and label them for destruction by phagocytes and lymphocytes (T and B)!
T lymphocytes / cells - Cell-mediated immunity, directly attacking infected cells.
B lymphocytes / cells - Humoral immunity, producing antibodies to neutralize pathogens.

Epitopes - They are protein structural features.

Q. How do you think the immune system changes as we age?
As we age, our immune system undergoes a number of changes that can affect its ability to protect us from infection and disease.

1 major change is the decrease in the production of certain types of white blood cells (such as T cells), which are important for fighting off infections. This decrease in cell production leads to an overall weakened immune system - making it more difficult for the body to fight off pathogens.

Ageing also affects the way our bodies respond to vaccinations. As we get older, our bodies become less responsive to vaccines, meaning they not be as effective at protecting us against diseases.

Finally, ageing can lead to increased inflammation throughout the body, which can further weaken the immune system and make it more susceptible to infection.

Q. How does the immune system change in response to HIV and AIDs?
Human Immunodeficiency Virus (HIV) is a virus that attacks and destroys the body’s T lymphocytes.
The reduction in the number of T lymphocytes in the body due to HIV can then lead to the development of Acquired Immune Deficiency Syndrome (AIDS).

Individuals with AIDS have a weakened immune system and so are more vulnerable to other infections such as tuberculosis and pneumonia.

There is no cure for HIV/AIDS although many scientists are trying to find one.
Currently, people who carry the HIV infection are given antiretroviral drugs.
These work by stopping the virus replicating in the body. This allows the immune system to repair itself and prevents the development of AIDS.

Q. What are the different types of antibodies and how do they work to fight off pathogens?
Antibodies are proteins produced by the immune system to help fight off pathogens.

In more scientific terms, antibodies neutralise pathogens and label them for destruction by phagocytes and lymphocytes (T and B)!
T lymphocytes / cells - Cell-mediated immunity, directly attacking infected cells.
B lymphocytes / cells - Humoral immunity, producing antibodies to neutralize pathogens.

There are 5 main types of antibodies: IgA, IgG, IgM, IgD and IgE.

IgA is found in mucosal secretions like saliva, tears and breast milk. It helps protect against viruses and bacteria that enter through the respiratory and digestive tracts.

IgG is the most abundant antibody type and it circulates throughout the body. It binds to antigens on the surface of invading pathogens and activates other components of the immune system to destroy them.

IgM is the 1st antibody to appear when a pathogen enters the body. It plays an important role in activating complement proteins which can damage or kill pathogens.

IgD is mainly found on B cells and helps regulate their activation.

Lastly, IgE is involved in allergic reaction. It binds to allergens and triggers the release of histamine which CAUSES inflammation > histamines boost blood flow in area of body affected by the allergen to help white blood cells travel to area and repair the tissues!

Q. What are the different types of white blood cells and their functions?
Yes, I am familiar with the different types of white blood cells. It is important to understand how each type of cell works in order to diagnose and treat diseases.

The 4 main types of white blood cells are: Neutrophils, eosinophils, basophils and lymphocytes.

Neutrophils are the most abundant type of white blood cell and they help fight off bacterial infections by engulfing and destroying bacteria.

Eosinophils help protect against parasites and allergies.

Basophils release histamine which helps to CAUSE inflammation.

Lymphocytes produce antibodies that help fight off viruses.

Q. How would you troubleshoot scientific equipment?

Identify the Issue: Observe any unusual sounds, displays, or performance issues.
Consult Manuals: Refer to the equipment’s user manual for specific troubleshooting steps.
Process of Elimination: Rule out the most obvious causes first and work systematically.
Seek Help: If the issue persists, consult with colleagues or contact the manufacturer for support.

E.g At fleet the Gyros machine gave split reps for one of the standard curves. I was required to carry out troubleshooting of the machine by performing needle washes and cleaning of the machine following the guidance in the machine protocol.

Q. That is the end of your interview, do you have any questions for the panel?

How do you protect staff from H+S implications such as the risk of infection from handling patient blood samples?
Can you tell me more about the team I would be working with?
What are the biggest challenges currently facing the department?
How does the NHS support professional development and career progression?
How do you see this role evolving over the next few years?

SCIENCE BASED QUESTIONS

Q. What is renal function and why is it important?
Renal Function - refers to how well your kidneys are working. The kidneys are responsible for several vital tasks in the body:

Filtering blood of waste products, regulating fluid balance, regulating electrolyte balance and acid-base balance, blood pressure regulation and red blood cell production (by producing erythropoietin which then stimulates RBC production).

Q. Why is acid base balance important?
Acid Base balance is crucial for maintaining the body’s homeostasis, which is the stable condition necessary for cells and organs to function properly. It is essential to maintain overall health and wellbeing.
Here are some key reasons it is important:

Enzyme function - an imbalance in pH can hinder enzyme reactions, affecting metabolism etc.

Cellular activities - cells rely on a stable pH to perform functions such as energy production, nutrient transport and waste removal.

Oxygen transport - Haemoglobin (protein in red blood cells that carries oxygen), is sensitive to pH levels. An imbalance can affect its ability to bind and release oxygen efficiently.

Electrolyte balance - acid base balance influences the levels of sodium, potassium and calcium which are ESSENTIAL for nerve function, muscle contraction and heart rhythm.

Nervous system function - is highly sensitive to changes in pH and an imbalance can lead to confusion, fatigue and even seizures.

Q. What is the Renin-Angiotensin System (RAS) and how does it function?

The RAS is a hormone system that helps regulate blood pressure and fluid balance in the body. Here is a simple summary:
1. Renin release - if blood pressure drops or sodium levels decrease, the kidneys release an enzyme called renin.

2. Angiotensin I Formation - Renin converts a protein called angiotensinogen produced by the liver into angiotensin I.
3. Angiotensin II Formation - Angiotensin I is then converted into angiotensin II by the angiotensin-converting enzyme (ACE) mainly in the lungs.

Effects of Angiotensin II:
Vasoconstriction - narrows blood vessels, increasing blood pressure.

Aldosterone release - simulates adrenal glands to release aldosterone which causes the kidneys to retain sodium and water, increasing blood volume and pressure.

ADH release - promotes the release of antidiuretic hormone (ADH) from the pituitary gland, which helps retain water.

Q. What are the roles of NADH and NAD+ in Glycolysis?
They play crucial roles in the energy extraction process from glucose. Without NAD+ and NADH, glycolysis would halt, and cells wouldn’t be able to efficiently extract energy from glucose.

1. NAD+ as an electron carrier - During glycolysis, it accepts electrons and hydrogen ions, becoming NADH.
2. Energy extraction - In one of the key steps in glycolysis, glyceraldehyde-3-phosphate is oxidised and NAD+ is reduced to NADH. This step is crucial for extracting high energy electrons from glucose.
3. ATP production - The electrons carried by NADH are later used in the electron transport chain (in aerobic respiration) to produce ATP, the cell’s main currency of energy.
4. Regeneration of NAD+ - For glycolysis to continue, NADH must be oxidised back to NAD+. This can happen through aerobic respiration or fermentation, depending on the presence of oxygen.

Q. What is Oxidative Phosphorylation and how does it work?
It is the FINAL step in cellular respiration, where the majority of ATP (energy) is produced. Steps:

1. Electron Transport Chain - Electrons from NADH and FADH2 are passed through a series of proteins in the inner mitochondrial membrane.
2. Proton Gradient - As electrons move through these proteins, protons H+ are pumped from the mitochondrial matrix to the intermembrane space, creating a gradient.
3. ATP Synthesis - Protons flow back into the matrix through ATP synthase, a protein that uses this flow to convert ADP into ATP.
4. Oxygen’s Role - Oxygen acts as the final electron acceptor, combining with electrons and protons to form water.

This process efficiently produces ATP which cells use for energy.

Q. Why is it important to maintain healthy levels of the electrolytes sodium and potassium in the body?
Essential for overall health and wellbeing.

Nerve Function - Both Na+ and K+ help generate and transmit electrical impulses in the nerves which are necessary for muscle contractions and communication between nerve cells.

Muscle Function - These electrolytes are vital for muscle contractions, including the heart muscle. Imbalances can lead to muscle weakness, cramps and even cardiac issues.

Fluid Balance - Na+ helps regulate the amount of water in and around cells. Proper balance helps prevent dehydration and ensures cells function optimally.

Blood Pressure Regulation - High Na+ levels can lead to hypertension, while adequate K+ intake can help to counteract sodium’s effects and lower blood pressure.

Acid Base Balance - Both electrolytes play a role in maintaining the body’s pH balance, which is crucial for enzyme function and overall metabolic processes.

Q. What are the main Thyroid hormones and what are their main functions?
The main thyroid hormones are Thyroxine (T4) and Triiodothyronine (T3). They play crucial roles in regulating metabolism, growth and development.

Functions:
Metabolism - they control the rate at which your body uses energy (calories), affecting weight gain or loss.
Heart Rate
Body Temperature
Digestive Function
Brain Development
Muscle Control

What happens if they go wrong?
Hypothyroidism - occurs when the thyroid gland DOESN’T produce enough hormones. Symptoms can include fatigue, weight gain and slowed heart rate.

Hyperthyroidism - when the thyroid gland produces TOO MUCH hormone. Symptoms include weight loss, rapid heartbeat, anxiety and tremors.

Goiter - An enlarged thyroid gland.

Thyroid nodules - lumps in the thyroid gland that can sometimes produce excess hormones or be cancerous.

Q. What is TSH and what is its role?
TSH is Thyroid Stimulating Hormone. It is produced by the pituitary gland in the brain and plays a crucial role in regulating thyroid gland function. Here is how it works:

1. Regulation - TSH stimulates the thyroid gland to produce and release the thyroid hormones T3 and T4.
2. Feedback Loop - When the thyroid hormone levels are LOW, the pituitary gland releases more TSH to stimulate the thyroid.
   Conversely, when thyroid hormone levels are HIGH, the pituitary gland reduces TSH production.

*3. Diagnostic Tool - Measuring TSH levels in the blood is a common way to assess thyroid function.
HIGH TSH levels can indicate Hypothyroidism (under-active thyroid) while LOW TSH levels can indicate Hyperthyroidism (overactive thyroid).

Maintaining proper TSH levels is essential for overall metabolic balance and health.

*Q. What is the Calprotectin Test? (On my clinical scientist shadowing day, I learnt that the STP students I spoke to had contributed to the development and validation of a calprotectin test at Frimley Park NHS in the POC department!)
Calprotectin Test - A faecal test used to detect inflammation in the intestines. Here is what it does:

Detects Inflammation - Calprotectin is a protein released by white blood cells in response to inflammation. High levels of calprotectin in your stool can indicate inflammation in the gastrointestinal tract.

Differentiates Conditions - It helps distinguish between inflammatory bowel disease (IBD) like Crohn’s Disease and ulcerative colitis and NON-inflammatory conditions like irritable bowel syndrome (IBS).

Monitors Disease Activity - For those with IBD, the test can monitor disease activity and predict flare ups!

Question 6

Haematology Rotation Example- what does coagulation entail?

Answer 6

Checking patient identifiers (?) and then you have to spin all the coagulation samples and check to make sure they put enough blood in the tube > have to have a 9:1 dilution of sodium citrate (the anti-coagulant to the blood) otherwise the tests don’t work.

Then just very simply loading them onto the analyser and run the test.

Observe people authorise the results - why some results might be high etc.

Question 7

Liver Function Tests

Answer 7

Liver Function Tests (LFTs) are a group of blood tests that provide valuable information about how your liver is functioning.

Alanine transaminase (ALT):
ALT is an enzyme primarily found in liver cells. Elevated ALT levels in the blood indicate hepatocellular injury, which can occur due to conditions like hepatitis, liver cirrhosis, drug-induced liver injury.
7-56 IU/L.

Aspartate aminotransferase (AST):
Similar to ALT, AST is also found in liver cells. Raised AST levels are another marker of hepatocellular injury. An AST:ALT ratio greater than 2:1 is suggestive of alcoholic liver disease.
11-47 IU/L

Alkaline phosphatase (ALP):
ALP is derived from biliary epithelial cells (lining the biliary tract) and bones. Elevated ALP levels can result from cholestasis (interrupted bile flow) due to gallstone disease, external compression of the biliary tract, or medication side effects.
30-120 IU/L

Gamma-glutamyltransferase (GGT):
GGT is present in hepatocytes (liver cells) and biliary epithelial cells. It’s a sensitive marker of liver damage and cholestasis, although it lacks specificity.

Bilirubin:
Bilirubin levels reflect liver function and can be direct (conjugated) or indirect (unconjugated). Elevated bilirubin may occur in conditions like cholestasis or liver disease.

Albumin:
Albumin is a protein produced by the liver. Low albumin levels may indicate liver dysfunction.

Reference ranges for these tests can vary between laboratories, so always check local guidelines.
Abnormal LFT results should prompt further investigation to determine the underlying cause.

Question 8

Cardiac Markers

Answer 8

Cardiac markers are substances released into the bloodstream when the heart is damaged or under stress. These biomarkers provide valuable information about heart health and can aid in diagnosing various cardiac conditions.

Troponin:
This is the most specific and sensitive marker for myocardial damage. Troponin is composed of three proteins: Troponin C, Cardiac Troponin I, and Cardiac Troponin T.
Troponin I, in particular, has a high affinity for detecting myocardial injury. It is released during a heart attack (myocardial infarction) from the cytosolic pool of myocytes. Troponin levels remain elevated for several days after an event.

Creatine Kinase (CK-MB):
CK-MB is relatively specific when skeletal muscle damage is not present. It is an enzyme released during heart injury. However, it is less specific than troponin.

High-Sensitivity Troponin:
A newer test that can measure very low levels of troponin. It provides increased specificity compared to CK-MB.

Myoglobin:
A protein that stores oxygen. It is released quickly but is not very specific. Myoglobin levels rise rapidly after heart damage.

These markers help diagnose and monitor heart-related conditions such as heart attacks, heart failure, and myocardial infarction.
By assessing the extent of cardiac damage, healthcare professionals can guide treatment decisions.

Question 9

What is the HbA1c test?

Answer 9

The HbA1c test, also known as the glycated hemoglobin test, measures your average blood sugar level over the past 2-3 months. It reflects how well your blood sugar has been controlled during that time. Here’s how it works:

Hemoglobin: A protein in red blood cells that carries oxygen.
Blood sugar (glucose): Provides energy to cells.
Glycation: Glucose binds to hemoglobin, forming HbA1c.

Test results:
Normal: HbA1c below 5.7%.

Prediabetes: HbA1c between 5.7% and 6.4%.

Diabetes: HbA1c of 6.5% or higher.
Target for diabetes management: Keep HbA1c levels below 7%.

Remember, HbA1c helps diagnose diabetes and assess glycemic control.
Regular testing is crucial for managing diabetes effectively.

Question 10

How is the Clinical Biochem STP programme structured?

Answer 10

The Scientist Training Programme (STP) in Clinical Biochemistry is a comprehensive training program designed to prepare aspiring Clinical Scientists for their roles in healthcare.

Key Components:

Core Modules, Rotation Modules, Specialist Modules.

Work-Based Learning:
Trainees engage in work-based learning and training within the NHS workplace. This practical component allows them to apply their academic and clinical knowledge and skills. Competencies and assessments are integral to work-based learning.

Academic Programme:
The STP combines an academic program (usually an MSc in Clinical Science) with the work-based component.
Understanding the core scientific principles!

Question 11

General methods used in Biochem

Answer 11

Automated analysis
Immunoassay
Ion selective electrode
Electrophoresis
Chromatography
Mass spectrometry
DNA technology
Quality assurance

Automated Analysis:
Clinical biochemistry laboratories employ highly sophisticated automated equipment to analyse a wide range of substances. For instance, tests that require large-scale measurements (such as potassium and urea levels in blood) are efficiently processed using these automated systems.

Immunoassay:
This technique detects and quantifies specific molecules (such as hormones, proteins, or drugs) by utilizing antibodies. Immunoassays are valuable for diagnosing various conditions and monitoring treatment progress.

Ion-selective electrode (ISE):
ISE is a remarkable transducer that converts changes in the concentration of a specific ion dissolved in a solution into an electrical potential.

Electrophoresis:
By separating charged molecules based on their mobility in an electric field, electrophoresis helps identify proteins, enzymes, and other biomolecules. It’s particularly useful for detecting abnormal protein patterns.

Chromatography:
Various types of chromatography are employed to separate and analyse complex mixtures. These methods allow precise identification of compounds.

Mass Spectrometry:
Mass spectrometry provides detailed information about the composition and structure of molecules. It’s especially valuable for detecting small molecules, metabolites, and drugs.

DNA Technology:
Clinical biochemistry increasingly relies on DNA-based techniques. Polymerase chain reaction (PCR), gene sequencing, and genetic testing play essential roles in diagnosing genetic disorders and assessing disease risk.

Quality Assurance:
Ensuring the accuracy and reliability of test results is critical. Clinical biochemists participate in quality control measures, validate new tests, and maintain high standards in laboratory practice.

Consultation and Interpretation:
Clinical biochemists collaborate with clinicians and general practitioners. They advise on appropriate test selection, interpret results, and recommend follow-up investigations when necessary.

Question 12

NHS Values

Answer 12

The NHS (National Health Service), which provides healthcare services in the United Kingdom, upholds six core values:

Working together for patients
Respect and dignity
Commitment to quality of care
Compassion
Improving lives
Everyone counts

Working Together for Patients:
Patients are the top priority, and all efforts are directed toward their well-being and care.

Respect and Dignity:
Every individual, whether a patient, family member, or staff, is valued. Their aspirations, commitments, and unique circumstances are respected and understood.

Commitment to Quality of Care:
The NHS insists on high-quality care, focusing on safety, effectiveness, and patient experience.

Compassion:
Compassion is central to the care provided. Staff respond with humanity and kindness to each person’s pain, distress, anxiety, or needs.

Improving Lives:
The NHS strives to enhance health, well-being, and people’s experiences within the healthcare system.

Everyone Counts:
Resources are maximized for the benefit of the entire community, ensuring that no one is excluded, discriminated against, or left behind.

Question 13

NHS Constitution

Answer 13

HIGH QUALITY HEALTHCARE AVAILABLE FOR ALL

Places patients at the heart of everything.
Collaboration.
NHS remains accountable to the public and patients it serves.

For patients and those using NHS services, the NHS Constitution emphasizes the following principles:

The NHS provides comprehensive services available to all based on clinical need, not an individual’s ability to pay.

Aspiring to excellence and professionalism, the NHS places patients at the heart of everything it does.

Collaboration across organizational boundaries and partnerships with other entities serve the interests of patients, local communities, and the wider population.

The NHS remains accountable to the public, communities, and the patients it serves.

*These values shape the NHS’s commitment to delivering high-quality healthcare and ensuring equitable access for all.

Question 14

Biggest hazard in the NHS - handling human blood samples

Answer 14

Risk of infection from handling patient blood samples > e.g. HIV, hepatitis etc. Could accidentally transmit virus by touching needle.

Health and safety precautions > at Fleet had to sign off SOP / documents before handling human samples and was warned about H+S implications.

What measures do you have in place to protect staff from potential infections when handling blood samples?

Certified samples vs uncertified:
Certified = screened for viruses such as hepatitis and HIV before use = safer.
Uncertified = not screened before use = not as safe.

Question 15

Give an example of working in a high pressure environment?

Important > links between departments

Answer 15

At Fleet I had to learn to juggle my tasks between both chemistry and assay departments. Good time management and communication with my placement supervisor was essential to balance the high workload and manage my stress in the high pressured lab environment.

E.g. STP > Clinical Biochem will provide samples to haematology and haematology will provide samples to immunology etc.

The links between departments are also key to provide high quality patient care.

Question 16

Good Scientific Practice

Answer 16

Purpose:
Patients are the number one priority and their safety in all settings where the healthcare science workforce
operate is paramount.
The purpose of Good Scientific Practice (GSP) is to set out the professional standards on which safe and good working practice is founded for all those in the healthcare science workforce.

The standards are essential for all members of the workforce to perform their job role activities; provide safe, effective patient care; and demonstrate their professionalism.

Domain 1: Professional practice – Patient-centred care; Scope of practice; Communication; Professional responsibilities; Working with others.
e.g. You put patients first and act in the interests of patients’ safety and well-being at all times.

Domain 2: Scientific practice – Data and reporting; Technical proficiency; Quality assurance.
e.g. Adhering to up-to-date standard operating procedures, competence with using equipment, engaging with health safety protocols, applying quality standards and seeking quality improvement also.

Domain 3: Clinical practice – Clinical activity; Clinical investigation and therapeutics
e.g. You obtain informed consent from individual parties (patients, or their carer) before you do the following: - - -
Undertake an investigation or examination.
Provide treatment.

Domain 4: Research, development and innovation – Research activity; Service development.
e.g. You act with honesty, probity and integrity in all stages of the research process, including by adhering to research governance frameworks and protocols.

Domain 5: Clinical leadership – Developing self; Leading others.
e.g. You demonstrate self-awareness, including about your leadership style and its impact on others.
You value and recognise your colleagues’ knowledge, skills and contribution to service delivery and patient care.
Celebrating team success.
Acting as a role model and leading by example and recognising when development is needed.

Question 17

What constitutes a good leader?

Answer 17

Self awareness

Treating people with respect

Good communication and friendly / approachable nature

Actively listening and providing feedback

Value your colleague’s knowledge and skills and address when development is needed.

Celebrating team success.

Being open to new ideas / having a growth mindset

Acting as a role model and leading by example.

Inspiring a positive work ethic / environment.

Question 18

Dealing with conflict example

Answer 18

Buffer thrown down the sink > resilience > able to bounce back when faced with adversity

Question 19

Giving or receiving feedback

Answer 19

Presentations at Fleet > received constructive feedback

When I’ve made mistakes at Fleet > learnt and acted on it.

Question 20

Using reflection to improve your practice/skills

Answer 20

Gyros project > made a few errors but learnt from this / reflected and did not repeat the mistakes.

Question 21

Clinical Biochem - what is expected of me?
LEARN

Answer 21

Patients that come into a hospital may have a blood sample taken, and some time later they’ll get their results back, with little knowledge about the processes that have occurred between these points in order to produce these result!

But the steps in between depend upon the work of clinical biochemists and a wide range of laboratory staff, each having a crucial role in the analysis of a sample.

Biochemical analysis has been crucial in the detection and treatment of diseases, including diabetes, inherited disorders/diseases, deficiencies, cancers, and many other illnesses.

In addition to routine work, a clinical biochemist also has a significant role in the development and validation of new tests being introduced in the department.
This ensures that the services available in the laboratory are of the highest standard and will ensure the most accurate clinical outcomes to the patient.

A clinical biochemist may also partake in research in the department.

It is important to know that STP clinical biochemistry specialism is more lab based and therefore has less patient interaction in comparison to other STP specialities!

Question 22

What does a day look like as a trainee biochemist?
LEARN

Answer 22

Currently most of the day-to-day work revolves around learning competencies, while also juggling learning the university content.

Some of the competencies I’ve worked towards so far include receiving, labelling, and storing samples, and more recently moving onto analysing the samples myself, using a wide range of techniques such as flow cytometry, mass spectrometry, and many more.

It’s also important to learn about how these analysers work to help with trouble shooting and maintenance, and how to read and report the results of the test.

Example - different types of blood samples:

Different samples require blood to be stored differently depending on the test. This is indicated with the different colour caps.

The purple tube can be used for a full blood count, measuring the amount of red blood cells, white blood cells and platelets that are present in the blood.

The green tube is for serum samples, that allows us to measure analytes like urea and electrolytes.

While the blue tube is used to screen the coagulation of a blood sample (the colours may vary in different hospitals!).

These tests allow us to pick up abnormalities in a patient’s blood sample, and can assist with the diagnosis of a wide range of diseases.

This has a big impact on patient outcomes, making the work of a biochemist vital for the NHS.

Question 23

What diseases are key to the clinical biochem speciality?

Answer 23

Liver diseases
Kidney disorders
Cardiovascular diseases (heart)
Diabetes mellitus
Cancer

Inherited metabolic disorders e.g. PKU - Phenylketonuria > a rare inherited disorder that affects how the body breaks down protein. People with PKU cannot break down the amino acid phenylalanine, which then builds up in their blood and brain. This can lead to brain damage, seizures and disabilities.

Hypertension (high blood pressure)
Obesity > + related health issues

Accurate diagnosis, treatment and monitoring relies on biochemical tests.

Question 24

*AKTA purifier Columns / Chromatography

Answer 24

Allthese purification approaches can be carried out using an AKTA FPLC ( Fast Protein Liquid Chromatography) system.

Comparable to HPLC (high performance liquid chromatography) butdesigned tooperateat lower back-pressures suitable for protein purification and characterisation.

How to clean and purify AKTA column before use:

3 chemicals are used for cleaning:
Sodium Hydroxide to remove microbes as it has anti-microbial properties.
Guanidine / Azide to denature/ remove any contaminating proteins or bugs and a buffer for extra cleaning.

It runs for 3 hours and the conductivity line will reach a really high peak as NaOH is a very strong conductor and then should fall back down and stay level.

Biomolecules are purified using chromatography techniques that separate them according to their specific properties. For example, size exclusion chromatography (SEC) is according to their size. High MW elutes off first, low MW last.

After use, good practice to wash AKTA pumps into water and leave over ethanol – will last longer.
Good AKTA practices workshop – training and development

I learnt how to use the AKTA at Fleet for protein purification:

The alarm pressure should always be set to 0.8 before you start.
We collected about 45 fractions in 12mm tubes. If using 12mm tubes > use the purple racks!
May use a superloop to inject sample onto column.

Cleaning the AKTA after use:

First remove the super loop and tubes and put away.
Put pump A and pump B into deionised water. Use a 10ml syringe to empty each of the 4 knobs and make sure there’s no air bubbles> make sure they are tight afterwards.
Then run pump wash basic until it levels off.
Put pump A and pump b into 100% ethanol and do the same thing > 10ml syringe to empty the knobs > tighten and run pump wash basic.
The AKTA will now sit in the 100% ethanol for the next person to use it.

Question 25

How would you deal with a discrepancy in your lab data?

Answer 25

If I noticed a discrepancy in my data, the first thing I would do is take a step back and assess the situation.
I would review all of the data to determine what caused the discrepancy and if it was an isolated incident or part of a larger pattern.
Once I had identified the source of the discrepancy, I would then work to find a solution. Depending on the nature of the issue, this could involve re-running experiments, revising protocols / MBRs, or further analysing the data.

During my time on placement, I have encountered a few data discrepancies and learnt how to identify and resolve them.

EXAMPLE > Gyros project: Testing of a protein-drug analyte for a biopharmaceutical company. Encountered a few errors when working on this project e.g. one assay run did not generate a standard curve, perhaps due to errors in making up the standards on my apart due to insufficient training and pipette inaccuracy.
To overcome this, I requested more training on correct pipette technique for tiny volumes and ran the assay again, taking extra care this time and thankfully, the second run worked! Important to request extra training when unsure, don’t be scared to ask and always pay attention to detail.

My attention to detail and problem-solving skills make me well-suited for this role.

Question 26

How would you deal with a difficult patient?

Answer 26

Dealing with difficult patients can be challenging, but it’s an essential part of providing quality healthcare:

Stay calm and professional at all times.

Listen to what the patient is saying > empathise and understand.

Put yourself in their shoes to try and understand their point of view.

Communicate effectively.

Assess the situation > self reflect and don’t let personal bias or stressors come into the equation!

Question 27

Things I Learnt at Fleet Bioprocessing

Answer 27

GENERAL

Immunoassays - Use antigen antibody recognition to detect and quantify an antigen in a sample: ‘Testing of an analyte to get a dose response’. ELISAs are the most common, examples include pregnancy and SARS-CoV-2 spike tests for COVID-19.
E.g. At Fleet:
Gyros FRD11 project > Assay layout was Streptavidin coated nanolitre microfluidic channels in the Bioaffy CDs > biotinylated capture antibody > analyte > detection antibody conjugated with Alexa Fluor > signal generation.

This method enhances sensitivity and allows for accurate detection of low-abundance targets in the sample. Amplifies signal output via streptavidin/biotin interaction and reduces passive adsorption.

Bioconjugation - Bioconjugation is a chemical technique used to couple two molecules together via a stable linkage, where at least one is a biomolecule. A Biomolecule is labelled to produce a conjugate.
E.g. Fleet developed a panel of monoclonal antibody-alkaline phosphatase conjugates for use in a novel TB test. Full robustness studies were executed by Fleet to confirm that all key variables were under control

What is the purpose of bioconjugation?
Bioconjugation enhances the functionality and specificity of biomolecules, making it a crucial tool in modern biomedical research! e.g. for use in assays, targeted drug delivery , diagnostics to detect the presence of biomarkers and in therapeutics to develop antibody-drug conjugates.

How to use and clean an AKTA column for purification > 3 chemicals are used for cleaning: Sodium Hydroxide to remove microbes as it has anti-microbial properties.
Guanidine to denature/ remove any contaminating proteins and a buffer for extra cleaning.
It runs for 3 hours and the conductivity line will reach a really high peak as NaOH is a very strong conductor and then should fall back down and stay level.

Size Exclusion Chromatography (SEC) on AKTA Columns:
Biomolecules are purified using chromatography techniques that separate them according to their specific properties. For example, SEC is according to their size. High MW elutes off first, low MW last.
Applications:
Protein Purification: SEC separates proteins based on size. Large heavy proteins (e.g., antibodies or conjugates) elute off first, while smaller ones (peptides) elute later.
Desalting and Buffer Exchange: SEC can also be used for these purposes.

OR Affinity Chromatography:
Where the conjugate will bind to the column and the junk will flow through the column. Then you switch the column into a different buffer to elute the purified product off the column and collect it!

Using Gyros machine > FRD job > sample prep, authorising and analysing results, evaluating and programming machine

Buffer reformulation project

How to use and clean a Freeze Dryer / lyophilisation > exothermic: ‘cake’ forms and temp drops further as water vapour sublimes off. After use back fill chamber with nitrogen and clean. Lyophilising is a way of storing proteins long term for stability, to prevent them denaturing > can be stored at 2-8 degrees.

Making regular buffers and recording on ‘Buffer Formulation Index Spreadsheet’ > including buffer 3, 32, 33, 99 and 115. SOP buffer sheets are then given for QC. Buffers resist changes in pH > can help to stabilise proteins and dilute substances to make them easier to work with.

Dilutions
Serial dilutions
Making IPA and Decon
Buffer reformulation project
Weekly fire alarm testing
Bioburden - excel
Lab coat change
Record keeping (lab book write ups, SOP buffer sheets, lab maintenance folder, LLRs)
Cleaning + calibrations of kit
Keeping paper work up to date
How to follow an SOP / MBR / LLR.
Shipping products at RT, with gel packs or with dry ice and contacting customers.
How to write a lab book write up correctly.
How to heat seal things.

SKILLS

Prioritising tasks
Attention to detail
Willingness to learn
Being flexible and adaptable when jobs are sprung upon me with little warning > working well under pressure.
Agile learning
Working to deadlines / under time pressure
Self - motivated
Collaboration / communication with other team members
The move to OneDrive - how this impacts a company
A chemistry director going on leave for 6 weeks > how a comings adapts to a key member of the team leaving > have to be agile / adaptable.

EQUIPMENT
* Freeze dryer
* Personal pipettes and digital repeat pipette
* Reverse Pipetting
* How to use and set up the chromatography separating column (AKTA) + super loop
* Plate reader and plate washer
* Gyros machine
* Optilite machine
* AKTA columns (protein purification).
* Different types of filters (Nalgene, bottle top, sartobran, minisart)
* Vortex mixers, roller mixers, incubators, balances, centrifuge
* Spectrophotometer (path length = 1cm)
* Nanodrop spectrophotometer (path length = 0.1cm, useful for very concentrated samples)
* Ultrasonic bath
* PD10 columns
* Integrity tester to check accuracy of filters

GENERAL STEPS TO MAKING A CONJUGATE
e.g. anti-malaria antibody - biotin conjugate
1.Addition of the two reagents > must be in the correct buffer to begin with! Conjugation using Biotin-XX-NHS > uses lysine chemistry > stable amide bond formed via NH2 groups.
2. Roller mixing / incubating
3. Desalting (buffer exchange / size exclusion chromatography) on PD10 columns (small scale) or AKTAs (large scale / purification ). In SEC high MW elutes off first aka. the conjugate product. Small MW is left behind and gets stuck in the pores.
4. Quench reaction with glycine
5. Measure UV to find concentration by using Beer-Lamberts law.
6. Find the biotin concentration via a HABA-Avidin assay and use the ratio of the concentrations to find out the biotin incorporation.
7. Dilute down to the concentration the customer wants using buffer.
8. Add preservatives such as Proclin 950 to kill bugs.
9. Filter into sterile bottles, label and store ready for shipment.

JOBS / SCIENCE STUFF

1. Gyros - Optimising a series of ‘fit for purpose’ assays for the quantification of a cancer and heart disease biomarker analyte in human serum. A series of parameters were validated:

Selectivity / sensitivity - Validating the selectivity and sensitivity of all the possible capture and detection antibody combinations as well as optimal concentrations to ensure they bind with high affinity and minimal cross reactivity + background noise.

The main point of Selectivity is to make sure that all samples can be distinguished from the blank (serum of same index @ same dilution eg 40%) to make sure that you`re not getting false positives or negatives!

Upper and lower LOQ determination - Important in determining the sensitivity and accuracy. The lower end is a good indicator of background level, and the higher end reflects capture antibody binding capacity. Results gave CVs of less than 20% = good precision.

Accuracy - QC samples to validate day to day performance and accuracy of the immunoassay. CV was below 15% = results are consistent and reliable over time.
Implementation of quality controls is critical to monitor assay performance and to detect data deviations, allowing for quick resolutions, therefore ensuring high quality, reproducible assay results.

Optimising the Gyrolab method - to use a slow spin method, 0.5%, for the analyte and detection.
This enabled more capture column interaction time with the aim of enhancing signal generation and compensating for low avidity of antibody.

Precision - Titration experiments determined the optimal concentrations of capture and detection antibodies. Sufficient S/N results suggests that there was minimal non-specific binding and good precision.
Determining the optimal dilution buffer to ensure ensure sample preservation and minimise non-specific binding.

Robustness - MRD was determined as 40% human serum tolerance. This ensures that measurements are reliable at low concentrations of analyte. Improves precision, accuracy, and dynamic range.

Gyros machine > streptavidin coated CD based technology with nanolitre microfluidic channels > useful for sandwich ELISAs with biotinylated capture antibody. Faster results, smaller reagent volumes + minimises human error by automated technology. Increases data quality, sensitivity and accuracy.

Optimisation of assay was successful > fulfilled all customer requirements for use in pre-clinical trials.

*2. Pepsin digest of HIV Ab (large scale)
Customer provides antibody: this specific one was very expensive as it was an Ab specific for HIV so critical that job works properly. Aim is to cleave off the Fc region of Ab by a pepsin digest leaving the two Fab regions joined by a disulphide bond > (Fab’)2.

Advantage of this is lower background noise meaning a more accurate assay using the HIV Ab which is VITAL for a blood test testing for HIV as a false positive or false negative result could have impact the treatment / diagnosis of the patient = effects their quality of life!

Job is carried out on AKTA column: desalt the Ab on column first > pepsin digest in citric acid pH 3.5 buffer (optimal balance for acidity of pepsin but to not kill Ab) then quench in pH 9 buffer to neutralise and finally purify columns in NaOH and then Azide buffer to kill bugs.

*Other example of a pepsin digest that Fleet do is for a Troponin Ab! Also very important that job is done correctly as Troponin is a cardiac marker > more sensitive and specific than CK MB. Vital that the assay works correctly and accurately so patients are not misdiagnosed and can be treated quickly. More critical that these tests work properly as opposed to a pregnancy test etc. It is essential to ensure accurate clinical outcomes.

4. Mouse IgG Ab Pepsin digest and biotinylation:
   First desalt into correct buffer for digest > pH 3.5 citric acid buffer. Then carry out pepsin digest to cleave off the Fc region, leaving just (Fab’)2.

Next, split (Fab’)2 into two Fab regions by breaking / reducing the disulphide bonds using TCEP > reduction.

Then you can conjugate each Fab region with biotin in 2 different ways.

1. Fab- Biotin - Maleimide
   This is Cysteine chemistry > conjugation via cystine residues > Maleimide Reagent reacts with Sulfhydryl on protein to = stable conjugate with biotin (THIOETHER BOND).
   Addition reaction = Simple.
   Thioether bond (stable) = pH 7-9, good site specificity. Ellman’s assay to monitor progress > it measures Sulphur.
   Buffer 99.
2. Fab- Biotin XX-NHS
   This is Lysine chemistry > NHS Ester reagent will react with lysine residues on primary amine of protein to = STABLE CONJUGATE (amide bond). Condensation reaction.
   Buffer 3.

Finally, biotin incorporation can be worked out by running a HABA-Avidin assay > Biotin displaces HABA as avidin has a high affinity for biotin > The HABA-avidin complex absorbs light at 500 nm > As Biotin displaces HABA, the absorption at 500 nm decreases proportionately.
The change in absorbance relates to the amount of biotin in the sample.

Customer can now use Mouse IgG Conjugated to Biotin > No Fc region so less background noise = better accuracy and precision. Biotin is useful for assays due to the streptavidin-biotin interaction > enables signal amplification, reduces non specific binding and more sensitive detection of analyte = more accurate and trustworthy results!

4. BCA assay > to calculate protein concentration / incorporation using BSA standards. Follow a protocol to generate a ‘quadratic polynomial’ calibration curve. R squared should = 1 for an accurate curve.
   *Responds to whether protein is conjugated / present or not > will turn purple if protein is present > the darker the purple colour = more protein present!

How to: Make up standards using water + bovine serum albumin protein standard, make up BCA working reagent, dispense 20µl of standards and test samples (protein /Ab) onto plate with 200µl of working reagent, incubate in 37 degree incubator for 30 mins then read @570nm to find unknown protein concentrations in plate reader by comparing to standard curve.

E.g. Fleet example > BCA Assay for GEM5 Arginine and Glycerol samples to determine protein concentration in conjugate. However, this was troubleshooting work, as first BCA assay did not work, so more testing was carried out to determine whether arginine was interfering with assay.
BSA standards were made up as normal, test samples on plate included: GEM5 arginine and glycerol samples, Arginine TBS buffer and 10% Glycerol TBS buffer as blanks. But two final positive control solutions were added to the assay which included a 0.5mg/ml BSA in arginine buffer and a 0.5mg/ml BSA in glycerol buffer. This is because we know BSA will work in the assay and will provide a known result, if arginine is interfering with the assay than the result will be different than expected.
Result > BCA assays do not work with arginine!

Theory: Redox reaction. Cu2+ reacts with Ab backbone.
Cu2+ (green) > Copper-BCA complex (purple, proportional to concentration of protein).
Best thing you can do is have protein with known concentration as a standard! (correction factors).

5. HABA-Avidin assay > for biotin incorporation .
   HABA > aromatic, delocalised.
   Biotin will displace HABA.
   Biotin displaces HABA as avidin has a high affinity for biotin > The HABA-avidin complex absorbs light at 500 nm > As Biotin displaces HABA, the absorption at 500 nm decreases proportionately.
   The change in absorbance relates to the amount of biotin in the sample.
   Less orange colour = more biotin bound and less HABA present.
6. Ellman’s Assay > Used for quantification of free sulfhydryl groups in solution.
   A solution of this compound produces a yellow-coloured product when it reacts with sulfhydryl groups. This yellow-coloured product indicates the presence of sulfhydryl groups and can be quantified using a spectrophotometer based on its strong absorbance @ 412nm.
   Therefore useful for Cysteine, Maleimide conjugations.

AND Reverse Ellman’s Assay testing to determine the concentration of maleimide groups. Cysteine standards are used to generate the standard curve, Ellman’s reagent used in the assay.
E.g. Fleet example > Reverse Ellman’s assay to test variation between the maleimide incorporations between new and old TAT-Mal samples. NEM was used as a positive control as it should give a maleimide incorporation of 1.0. Both the new and old TAT-Mal gave incorporations of around 0.5, both lower than expected (the closer to 1 the better), but consistent results.

7. TNBS Assay > Rapid and sensitive method used to determine the concentration of free amino groups in a sample.

Assay that tells you how many amines on protein you are measuring (NH2) groups / amine concentration > can use to work out the incorporation / if conjugation has worked.
It uses a standard curve with glycine solutions > as glycine is the simplest amino acid with one amine group so can use this as a comparison to the test sample to calculate the number of amines on protein.
Result = how many amines there are on molecule / conjugate.

Can also use to work out if conjugation has worked by comparing amine concentration of protein vs conjugate:
If amine incorporation decreases for the protein after TNBS assay but increases for the conjugate, this suggests that conjugation has been successful, as it means that conjugation via the amine groups has happened.
For a conjugations that work via amines > i.e. lysine residues in proteins.

Procedure:
Dissolve the sample (protein or small molecules) in a reaction buffer.
Add a solution of 0.01% TNBSA to the sample.
Incubate the mixture at 37°C for two hours.
Add 10% SDS and 1 N HCl to stop the reaction.
Measure the absorbance of the solution at 335 nm.
Quantitative determination can be done by comparing to a standard curve generated using an amine-containing compound (e.g., amino acid - Glycine) under identical conditions.

*8. Biotin : BSA (large scale)
Conjugate is made with BSA and Biotin-XX-NHS ready for purification on the AKTA.
AKTA is desalted into correct buffer to begin with so it is ready for purification of conjugate > check that this has happened by equilibrating the AKTA overnight and look at the trace > first buffer to desalt into is Azide to kill contaminating bugs / microbes, this has a low conductivity. Then desalt into PBS buffer as it is the right pH for the conjugate and has a high conductivity. Will know is desalt has happened because AKTA trace will go from low - high conductivity.

This is carried out on the AKTAs using large columns with size-exclusion chromatography (SEC). First, the Biotin:BSA conjugate sample is injected onto the column with a P6000 pump. Sample will slowly move down the column as it is purified via SEC > high MW molecules like the Biotin : BSA conjugate won’t be retained in the pores so fall through very quickly and come off first. Low MW molecules that are impure / unreacted such as unreacted Biotin-XX moves through slower as they get trapped in the pores so come off last.
High MW pure product = comes off first.
Low MW = comes off last.
So, the first peak seen on the AKTA trace is the higher MW Biotin:BSA product which is now a pure conjugate and can be fraction collected. Subsequent peaks are lower MW impurities / unreacted molecules > do NOT want to collect these!

Next, the product is filtered lots more times to ensure there are no impurities and Proclin 950 is added as a preservative so the conjugate can be stored without any contamination as BSA is prone to bugs growing.

Filter is tested with integrity tester after use to prove that it is filtering accurately.

Then, a HABA-Avidin assay is carried out to determine the biotin incorporation of the conjugate > should be the same every time as the method is the same every time = quality. The customer asks for a specific biotin incorporation.
Biotin:BSA sample is diluted first to 0.2mg/ml in PBS buffer for an accurate measurement > colour should still be orange to prove that some HABA is still bound > if colour turns yellow this means there is no HABA left and you have gone too far and all the HABA binding sites are saturated with biotin! Not good as now you won’t be able to measure the biotin incorporation.

Finally, the AKTA is cleaned with Sodium Hydroxide to remove anything that is non-specifically bound to the column matrix and left over buffered Azide to prevent bugs growing.

What is the chemistry happening here?
Lysine chemistry. The Biotin-XX-NHS reagent will react with the lysine residues on the BSA protein side chain. This lysine residue (NH2) acts as a nucleophile by donating a pair of electrons to the Biotin-XX-NHS reagent, causing the NHS group to fall off and leading to the formation of a STABLE AMIDE BOND.

9. ELISA Assay
   ELISA is based on the principle of antigen-antibody binding and enzyme-mediated colour change or fluorescence to generate a signal. Example I carried out at Fleet was a sandwich ELISA to evaluate precision:

Remember that sandwich ELISAs measure antigens between two layers of antibodies (capture and detection antibodies).

• Purpose : Precision investigation -> Multiple runs with multiple operators to account for variability in normal conditions.
  -> QC samples to look at the precision of the assay by reading the concentrations (absorbance read) off the curve produced by the assay.

Plate layout:
- Streptavidin coated plate
- Biotinylated TNFa (capture)
- Bispecific Antibody (Antigen – being measured)
- IL23 – HRP (detection)
- Sureblue (TMB) – Activates HRP - blue colour change
- HCl – Deactivates HRP - yellow colour change
- Measure absorbance @ 450nm
- Colorimetric reaction

Results:
- Generate standard curve using MyCurveFit of normal conditions vs QC samples for both operators for comparison. Normal: curve generated from concentrations and average responses. All %CVs were below 20% = acceptable. QCs: compare to previous calibration curve to calculate unknown concentrations. %CV all lower than 10% = acceptable. % difference should also be minimal as standard concentrations vs QC concentrations should be simialr.
%CV results were similar for both operators and curves all looked similar, suggesting adequate precision.

Method:
1 in 2 dilutions.

Prepare the Surface (Coating with Capture Antibody). Coat the wells of a PVC microtiter plate with the capture antibody in carbonate/bicarbonate buffer.
- Incubate.
- Wash the plate twice with 200 μL PBS buffer.
Blocking and Adding Samples. Block the remaining protein-binding sites in the coated wells by adding blocking buffer.
- Incubate.
- Wash plate with PBS buffer.
- Add 100 μL of diluted samples to each well. Always compare the signal of unknown samples against those of a standard curve.
Run standards in triplicates and blank with each plate.
- Incubate for 90 minutes at 37°C.
Incubation with Detection and Secondary Antibody
- Add diluted detection antibody to each well.
-Incubate
-Wash the plate to remove unbound antibodies.
-Follow up with the addition of a secondary antibody (labelled with an enzyme or other detection system).
-Incubate again.
-Wash the plate to remove any unbound secondary antibodies.
- Add Sureblue (TMB), start timer for 10 mins.
- Add HCL to stop reaction and measure absorbance. TMB produces the colorimetric signal that is read by the ELISA plate reader.

-Ensure that the concentration of standards spans the most dynamic detection range of antibody binding. You may need to optimize the concentration range to obtain a suitable standard curve.

10. SDS Page to test the purity of lots of different proteins accompanied by a Western Blot

A type of Gel electrophoresis. SDS is a denaturant that gives a uniform NEGATIVE charge to all proteins being measured in the gel > it separates proteins based on their molecular weight.
The proteins we tested were conjugates containing streptavidin.

Theory
Principle: SDS-PAGE separates proteins by their mass using an electric field. The method involves the use of sodium dodecyl sulfate (SDS), a detergent, and a polyacrylamide gel.

Polyacrylamide Gel: The gel acts as a sieve, allowing proteins to migrate through it. Slab gels (sandwiched between glass plates) are more common now.

SDS: SDS binds to proteins, masking their intrinsic charge and giving them similar charge-to-mass ratios. It unfolds both polar and nonpolar sections of protein structure.

Electrophoresis: When an electric field is applied, proteins migrate through the gel at different rates based on their mass. Smaller proteins move faster than larger ones.

Visualization: After separation, proteins can be stained to make them visible, allowing analysis of their molecular weights and quantities.

• We used SDS to check the purity of proteins.
• If protein is pure should only see 1 band. If lots of bands are seen = impure protein.
• Load samples, blank, standards, ladder and controls in the gel wells at the SAME concentrations > here was 0.6µg/ml > to allow for comparison > this is critical! Always load the same amount of sample in concentration.
• Standard ladder comprises of known protein molecular weights and therefore is used to find the MWs of proteins being run on the gel by comparison of bands. E.g. A band of a protein sample that is at equal level to the 20MW band of the standard ladder means the MW of that protein must be 20!
• Controls were streptavidin and BSA > BSA as a negative control as it does NOT bind biotin. Streptavidin binds strongly to biotin.
• The gel flow is from positive to negative charge and separation is based on mass as all the proteins are uniformly negatively charged. Higher up the gel = heavier MW and lower down the gel = lighter MW.
• Bands for the standards should all look consistent.
• All our proteins here are negatively charged so will move to the positive side > will always move to the side of opposite charge!

Western Blot - used to detect specific proteins in a sample.

• Final confirmation of protein purity > do after SDS page.
• Tag the protein being tested with something specific to the protein / Ab > it will bind and show as a band > to confirm its specificity.
• Used to confirm if the protein conjugate binds specifically to the Biotin-HRP tag as there is streptavidin in the conjugate.
• Cross check with an HRP only tag as a control > conjugate should not bind > it should only be specific for Biotin-HRP.

Principle
Separation by Size: Proteins are separated based on their size using gel electrophoresis.

Transfer to a Solid Support: The separated proteins are then transferred to a membrane (usually nitrocellulose or PVDF).

Antibody Labelling: An antibody is created to recognize and bind to a specific target protein > here want to see if Ab-streptavidin conjugate binds specifically to biotin-HRP.
The membrane is washed with a solution containing the primary antibody, and excess antibody is removed.
A secondary antibody is then added, which recognizes and binds to the primary antibody. T
he secondary antibody is visualized through various methods (such as staining) allowing detection of the specific target protein.

Purpose of blocking buffer and washing steps
The blocking buffer is a crucial step in western blotting, serving to prevent or reduce nonspecific binding of the primary and secondary antibodies to the membrane itself > reduces background interference.
E.g. Milk.

Proper washing ensures that nonspecific binding is minimized. It reduces background, optimises sensitivity and enhances signal.

Application
Used to detect the presence of a specific single protein within a complex mixture of proteins.

Results
Were successful! One western blot showed bands only for the biotin-HRP test samples and the other western blot showed no bands at all.
This is good as means our protein conjugate is specific only for biotin which was what we expected. The streptavidin binds to the biotin.

11. Following an MBR example as the main operator > ATL354/B
    Making a 16L scale of wash reagent for a customer. High attention to detail, concentration and methodical working key for success.
    Learnt how to use integrity tester.

*12. Optilite machine testing:
The Optilite machine is an advanced benchtop analyser, fully optimized and automated for high throughput and quality results in special protein testing, typically used in hospital labs to provide results from clinical sample analysis (blood, urine etc).

Example at Fleet - Validating Optilite machine reagents for DNaseB detection in human blood samples.

DNaseB is produced in response to Streptococcal infections > patients infected with this disease will produce antibodies against DNaseB > anti-DNaseB. Therefore, the optilite machine reagents used for DNaseB testing must contain the DNaseB antigen so it can bind to anti-DNaseB antibodies in the infected patient and confirm the disease diagnosis!

Fleet made Streptavidin Latex particles conjugated to DNaseB in house:
Latex beads have activated Carbonyl groups that bind to Streptavidin, Streptavidin then binds to biotin (high affinity interaction) and then the DNaseB antigen will bind to biotin to form the conjugate. This then forms the Optilite DNaseB reagent along with the addition of an optilite buffer.

These reagents will then be put in the machine ready for validation testing > the same samples are run three different times to ensure accuracy, all three curves should look the same (linear) = validation.
The optilite reagents are also run against DNaseB positive and negative calibration soltuions to see if the reagents produce FALSE positive or negative results > they shouldn’t if validation works correctly. With a clinical sample, it is important false positives and negatives are avoided to ensure the most accurate patient outcomes.

If the results of the DNaseB reagents look good, they will then be sent off to a regulation body such as FDA. If approved, the DNaseB optilite reagents can be used in hospital labs to detect streptococcal infections in clinical samples.

KEY POINT:
Vital that validation testing is carried out before DNaseB reagents are used in IVDs (In Vitro Diagnostic) medical devices or in pre-clinical testing in hospital labs for clinical samples from patients. Important that results are accurate, false positive and negative results are avoided so that the patient can have the most accurate outcome in terms of diagnosis and treatment.

Calibrating the plate washer with brilliant blue solution (part of quality assurance):
What I learnt from the plate washer calibration:

Tests check that the plate washer and reading are washing and reading correctly > any brilliant blue leftover will give a high reading > suggests contamination.
An important quality control measure and good lab practice > ensures a controlled environment.
Excel
Reverse pipetting to minimise air bubbles
Making up brilliant blue solution
Using a plate reader > importance of rinsing with DI water and wash buffer.
Excel and IT skills
Following SOPs
Updating calibration records
Attention to detail
Quality control

Question 28

Quality measures at Fleet

Answer 28

Dedicated buffer making bottles for BSA, separate filters, magnetic stirrers for BSA and separate washing up brush for BSA containing bottles > to avoid contamination.

Separate washing brushes for certain chemicals > eg. A BSA only brush.

Following MBRs and SOPs and using exact lot numbers and cat codes for raw materials. Sometimes customer specific materials too.

Line clearances and pH checks for SOPs and MBRs.

QCs of lab book write ups, SOPs and MBRs.

Testing of new batches of HRP before use > critical that quality of product is up to scratch > could impact results of jobs for customers.

Monthly calibrations / changing of lab coats / stock expiry date checks

Buffer expiry date checks > weekly

Monthly lab coat change

Storing washed buffer bottles with IPA

Recording all duties in lab maintenance folder

Monthly Bioburden settle plate data to ensure bacteria and fungi levels are below action limit > if over an extra lab clean is issued.

Monthly lab clean.

Monthly cleaning of Laminar flow cabinets.

Weekly fire alarm testing > health and safety

Wearing PPE

Question 29

Valuable things I’ve learnt

(Useful to mention in interviews / applications)
PRINT

Answer 29

• Laboratory experience
• Diversified experience working between 2 departments > immunoassay + Chem
• Experience in automated and manual immunoassay (ELISA) (Gyros)
• Troubleshooting / calibrating laboratory equipment
• Assay optimisation (FRD11)
• Quality management systems (QMS) experience > Audit and experience of quality control measures (NCNs etc and following SOPs and MBRs on a daily basis)
• Technical skills gained in lab techniques > understanding of analytical techniques.
• Knowledge of health + safety procedures (meetings, fire alarms, monthly check ups, PPE).
• Presentation skills > research ability + competence (fleet presentations and university).
• Teamwork, working well under pressure and in fast paced environment, resilience, communication.
• NHS motivation > ground breaking development opportunities and improving patients’ lives
• Learnt how to plan and prioritise your scientific workload on a day-to-day basis

Why I want to work in lab / for NHS?
* More than 70% patient healthcare decisions depends on the results of tests carried out in a laboratory.
* Crucial component to deliver high quality patient care
* NHS values
* Importance of accuracy, efficiency and attention to detail.
* Help to save lives and make a difference.
* Forefront of research + innovation = patients continually receive the very best healthcare.

Question 30

What are the different types of white blood cells and their functions?

Answer 30

Yes, I am familiar with the different types of white blood cells. It is important to understand how each type of cell works in order to diagnose and treat diseases.

The 4 main types of white blood cells are: Neutrophils, eosinophils, basophils and lymphocytes.

Neutrophils are the most abundant type of white blood cell and they help fight off bacterial infections by engulfing and destroying bacteria.

Eosinophils help protect against parasites and allergies.

Basophils release histamine which helps to CAUSE inflammation.

Lymphocytes produce antibodies that help fight off viruses.

Question 31

What are the different types of antibodies and how do they work to fight off pathogens?

Answer 31

Antibodies are proteins produced by the immune system to help fight off pathogens.

In more scientific terms, antibodies neutralise pathogens and label them for destruction by phagocytes and lymphocytes (T and B)!
T lymphocytes / cells - Cell-mediated immunity, directly attacking infected cells.
B lymphocytes / cells - Humoral immunity, producing antibodies to neutralize pathogens.

There are 5 main types of antibodies: IgA, IgG, IgM, IgD and IgE.

IgA is found in mucosal secretions like saliva, tears and breast milk. It helps protect against viruses and bacteria that enter through the respiratory and digestive tracts.

IgG is the most abundant antibody type and it circulates throughout the body. It binds to antigens on the surface of invading pathogens and activates other components of the immune system to destroy them.

IgM is the 1st antibody to appear when a pathogen enters the body. It plays an important role in activating complement proteins which can damage or kill pathogens.

IgD is mainly found on B cells and helps regulate their activation.

Lastly, IgE is involved in allergic reaction. It binds to allergens and triggers the release of histamine which CAUSES inflammation > histamines boost blood flow in area of body affected by the allergen to help white blood cells travel to area and repair the tissues!

Question 32

3. How do you think the immune system changes as we age?

Answer 32

As we age, our immune system undergoes a number of changes that can affect its ability to protect us from infection and disease.

1 major change is the decrease in the production of certain types of white blood cells (such as T cells), which are important for fighting off infections. This decrease in cell production leads to an overall weakened immune system - making it more difficult for the body to fight off pathogens.

Ageing also affects the way our bodies respond to vaccinations. As we get older, our bodies become less responsive to vaccines, meaning they not be as effective at protecting us against diseases.

Finally, ageing can lead to increased inflammation throughout the body, which can further weaken the immune system and make it more susceptible to infection.

Question 33

What are antibodies?

What are epitopes?

Answer 33

Antibodies are proteins produced by the immune system to help fight off pathogens.

In more scientific terms, antibodies neutralise pathogens and label them for destruction by phagocytes and lymphocytes (T and B)!
T lymphocytes / cells - Cell-mediated immunity, directly attacking infected cells.
B lymphocytes / cells - Humoral immunity, producing antibodies to neutralize pathogens.

Epitopes - They are protein structural features.

Question 34

How does the immune system change in response to HIV and AIDs?

Answer 34

Human Immunodeficiency Virus (HIV) is a virus that attacks and destroys the body’s T lymphocytes.
The reduction in the number of T lymphocytes in the body due to HIV can then lead to the development of Acquired Immune Deficiency Syndrome (AIDS).

Individuals with AIDS have a weakened immune system and so are more vulnerable to other infections such as tuberculosis and pneumonia.

There is no cure for HIV/AIDS although many scientists are trying to find one.
Currently, people who carry the HIV infection are given antiretroviral drugs.
These work by stopping the virus replicating in the body. This allows the immune system to repair itself and prevents the development of AIDS.

Question 35

How does the immune system change in response to allergies?

Answer 35

When a person who is allergic to a substance encounters it, the immune system may react by producing antibodies that “attack” the allergen.
This can cause wheezing, itching, runny nose, watery or itchy eyes, and other symptoms.

The immune system will produce immunoglobulin E, IgE, antibodies for each allergen.
The antibodies will cause cells in the body (mast cells + basophils) to produce histamine.

Once the allergen binds to IgE, specific types of cells (Mast Cells) will release chemicals that trigger the symptoms of the allergic reaction.

Question 36

What is an antigen?

Answer 36

The immune system can be activated by a lot of different things that the body doesn’t recognize as its own.

These are called antigens. Examples of antigens include the proteins on the surfaces of bacteria, fungi and viruses.

Question 37

What is an autoimmune response?

Answer 37

The body’s own cells have proteins on their surface, too. But those proteins don’t usually trigger the immune system to fight the cells.

Sometimes the immune system mistakenly thinks that the body’s own cells are foreign cells. It then attacks healthy, harmless cells in the body.
This is known as an autoimmune response.

Examples of autoimmune diseases: Crohn’s disease, Lupus, Type 1 diabetes.

Question 38

Streptavidin-biotin

Answer 38

Coating of antibodies/antigens to polystyrene plates by passive adsorption is very inefficient and can be detrimental to the protein

A common way to address this is to use streptavidin coated plates with biotinylated antibody or antigen:

Preserve antibody/antigen presentation
Reduce non-specific binding
Reduce usage of critical reagents
Amplification

Streptavidin is a 52 kDa protein (tetramer) purified from the bacterium Streptomyces avidinii.

Streptavidin has an extraordinarily high affinity for biotin (also known as vitamin B7 or vitamin H).

The binding of biotin to streptavidin is one of the strongest NON-covalent interactions known in nature, even stronger than the antibody/antigen reaction.

Each streptavidin monomer can bind one biotin molecule, allowing a streptavidin protein to maximally bind four biotins.

EXAMPLE OF SANDIWCH ELISA USING STREPTAVIDIN-BITOIN INTERACTION:

Gyros FRD11 project > Assay layout was Streptavidin coated nanolitre microfluidic channels in the Bioaffy CDs > biotinylated capture antibody > analyte > detection antibody conjugated with Alexa Fluor > signal generation.

This method enhances sensitivity and allows for accurate detection of low-abundance targets in the sample. Enhances signal output.

So, in summary, sandwich ELISA with streptavidin-biotin detection combines the high specificity of sandwich ELISAs with the signal amplification provided by streptavidin-biotin interactions, making it a powerful tool for detecting antigens!

Question 39

What is an ELISA?

Answer 39

An ELISA, or enzyme-linked immunosorbent assay, is a laboratory technique that can detect and measure the presence of specific molecules, such as proteins, antibodies, or hormones, in a liquid sample.

ELISA is based on the principle of antigen-antibody binding and enzyme-mediated colour change. ELISA can be used for various purposes, such as diagnosing diseases, testing for pregnancy, or monitoring immune responses.

Question 40

What is an immunoassay?

What is an assay?

What is a hapten?

Answer 40

Immunoassays use antigen antibody recognition to detect and quantify an analyte in a sample. Immunoassays are valuable for diagnosing various conditions and monitoring treatment progress.

ELISAs are the most common examples.

Assay - testing of an analyte to generate a dose response.

Hapten - Peptides and other small molecules that are used as antigens are referred to as haptens.

They are able to act as recognition sites for production of specific antibodies but cannot by themselves stimulate the necessary immune response.

Haptens can be made immunogenic by coupling them to a suitable carrier molecule.

Question 41

What is bioconjugation?

Answer 41

Bioconjugation is a chemical technique used to couple two molecules together via a stable linkage, where at least one is a biomolecule.

A Biomolecule is labelled to produce a conjugate.
Can have protein-protein conjugates
Or
Protein-small molecule conjugates etc

Examples of biomolecules:
Protein > A Protein biomolecule is made up of amino acids, arranged in a particular sequence (primary structure). Structure defines function. Enzyme, Nucleic acid, Carbohydrate.

Examples of biomolecules used at Fleet: Antibodies (IgG, IgM), Peroxidase from Horseradish (HRP), Alkaline Phosphatase (AP), Avidin / Streptavidin, BSA.

Examples of labels used at Fleet: Biotin, Fluorophores (small molecule, FTC.ED, AF), Enzymes (HRP, AP), Small molecule drugs / toxins, Haptens, Alexa Fluor.

Examples of conjugations at Fleet: Biotin-BSA, Ab-Biotin, Streptavidin-HRP, Ab- Alexa Fluor 488 (different numbers mean different absorbances = different colours), HRP-DES-Ab.

APPLICATIONS

Assay reagents.
Assay = Test of an analyte to get a dose response.

Immunology

Diagnostics (pregnancy test, thyroid function).

Veterinary

Therapeutics

Question 42

CV and Useful Application Bits

Answer 42

YEAR IN INDUSTRY PLACEMENT - SKILLS

Valuable for my professional development.
Accuracy, efficiency and attention to detail.
Laboratory experience
Diversified experience working between 2 departments > immunoassay + Chem
Experience in automated and manual immunoassay (ELISA) (Gyros)
Troubleshooting / calibrating laboratory equipment
Assay optimisation (FRD11)
Quality management systems (QMS) experience > Audit and experience of quality control measures (NCNs etc)
Technical skills gained in lab techniques > understanding of analytical techniques.
Knowledge of health + safety procedures (meetings, fire alarms, monthly check ups, PPE).
Presentation skills > research ability + competence (fleet presentations and university).

CV
Self-motivated and disciplined, I am a second-year Veterinary Biosciences student with a willingness to learn and a love for science-based approaches. With strong interpersonal skills and a common
sense approach gained through part time jobs and work experience, I am a great team player with high attention to detail.
My year long industrial placement in a specialist chemistry laboratory provided me with a range of practical, analytical and critical thinking skills, enhancing my professional development.
‘Working closely with our skilled team, they will gain hands-on industry experience, develop essential skills, and deepen their understanding of the life sciences industry.’
“I had a fantastic time during my placement at Fleet Bioprocessing. I developed invaluable professional and scientific skills, and it inspired me to pursue a career in clinical biochemistry.”
A motivated and CURIOUS Biomedical Scientist with laboratory, commercial-related roles and voluntary work experience. Reliable, trustworthy, hardworking, and eager to learn and constantly improve. Open-minded and seriously committed, seeking to enhance personal and professional knowledge through competitive work experience. Perseverance and determination are instrumental to personal development in addition to academic and professional life.

UNI

Bsc - Biosciences
Basic, applied and clinical aspects of biochemistry, immunology, genetics and pathology: lectures, seminars, practical skills, laboratory skills, tutorials, poster and paper presentations, individual and group projects.
Lectures and skill-based laboratories provided a theoretical and practical understanding of relevant techniques.
4th year research project:
Activities and societies: Ladies tennis first team, weekly dog walking, yoga, netball, weightlifting.

FLEET BIOPROCESSING LTD | Hartley Wintney, Hampshire | Sep 2023 - Sep 2024

Skills attained:
- Diversified bioconjugation and immunoassay laboratory experience

• Technical skills gained in lab techniques such as assay development and optimisation, ELISA, chromatography, protein purification, spectrophotometry including nanodrop, protein assays, buffer formulation (small and large scale), buffer titrations and reformulation, lyophilisation (freeze drying), SDS page, western blot and pipette accuracy.
• Experience in operating unique automated immunoassay equipment, including machine programming and evaluating results e.g. when undertaking my research project on immunoassay optimisation on the Gyrolab machine
• Knowledge of health + safety procedures and Quality Management Systems experience SUCH AS controlled documents, quality records and traceability!*
• Calibration and troubleshooting of scientific equipment
• Maintaining good laboratory practice e.g. lab book write ups, quick reviewing of work, line clearances, performing routine daily lab duties and updating equipment files
• Attending weekly team production meetings
• Understanding the pressures and pace of laboratory work
• Recognise and comprehend the impact work has on patients
• Good time management (when juggling tasks between two departments), Initiative, Good resilience and time management essential when working in a high pressure, fast paced lab environment, team working (working closely with other scientists / liaising with supervisor), communication and attention to detail, critical and analytical thinking developed by planning and performing own experiments (when carrying out own experiments, time pressure).
• Shadowing / assisting with senior scientist roles.
• Training of new staff

-Strong academic performance.

-Strong technical and laboratory skills.

-Strong critical thinking, communication and collaboration skills.

-Demonstrate a genuine passion for science and a strong interest in clinical science as a career.

PLACEMENT SUPERVISOR FEEDBACK

Transferrable Competencies: (rating B)

The new environment can be overwhelming for students. We saw that Olivia became progressively more confident in her role.

While some of the experiments performed did require specialist knowledge and experience, Olivia demonstrated that she could work through the logic and draw her own conclusions.

Application of Academic Learning: (rating B)

Initially, some of the work in the chemistry group was outside of Olivia’s comfort zone. However, over time she developed her knowledge base. For example, Olivia completed a number of buffer optimisation reactions which helps us improve our processes. She successfully performed “Reversed Ellman’s Assay” which as part of key troubleshooting activities. Where opportunities allowed, Olivia was able to perform assays using our Gyros equipment with limited supervision.

Professionalism: (rating A)

Olivia was approachable and was happy to help her colleagues.

We were pleased to see that the routine tasks assigned to her were completed in a timely fashion and recorded in accordance with our procedures. A number of these tasks would be considered unexciting but necessary. Nonetheless, Olivia completed them diligently.

Organisational awareness: (rating A)

During her placement, Olivia was provided with access to all part of our organisation including Finance and Quality as well as the Chemistry and Assay sections. Olivia attended the weekly planning meetings for the chemistry and assay teams.

The demands of working in a controlled environment can seem overwhelming at first. Quality and HSE guidelines need to be followed closely. Records need to be accurate and well maintained not just for the experimental details but also for the equipment and raw materials.

Olivia demonstrated that she was able to follow standard operating procedures to become a competent member of staff.

Olivia was present for several significant changes within this organisation and she showed eagerness to adopt new practices and circumstances.

PLACEMENT TUTOR FEEDBACK

In terms of your reflection (my feedback) I really thought that you approached the PDR form with great thought and attention. You were honest in your appraisal of your proficiencies, and you reflected on both the good work that you performed but also when the work did not go so well. I would like to congratulate you on doing such a good placement, which in some ways was outside of your background experience. Your capacity to embrace such a placement and to make the very most of it must be applauded. I am sure that you will use many of the learnings both professionally and at a subject level going forward.

Question 43

What is Point of Care Testing? (POCT)

Answer 43

Point of Care Testing (POCT) in the NHS refers to conducting diagnostic tests at or near the location of the patient, rather than in a conventional laboratory setting.

The goal of POCT is to collect specimens and obtain accurate results quickly, allowing for immediate clinical decisions!

These tests typically involve blood and urine testing.

The Point of Care Testing (POCT) team brings benefits to patients in allowing timely decision-making, often permitting life-saving interventions to be administered immediately = better care experience for patients!

Question 44

Current research in the NHS

Answer 44

Point of care testing team (POCT) in the NHS added a new blood test called C-reactive protein (CRP) >
CRP is a protein made by the liver and levels increase when there is inflammation or infection in the body > CRP is one of the best indicators of inflammation in the body!
CRP testing empowered timely decision-making for antibiotic prescriptions. Instead of waiting for lab results, the teams could make real-time decisions > prevented 13 admissions to hospital!

New higher sensitivity troponin test >
High-sensitivity troponin tests can detect lower levels of troponin T in the blood earlier than standard tests. They rule out of a specific type of heart attack called NSTEMI (non-ST-segment elevation myocardial infarction) earlier. Reduces hospital admissions!

Antimicrobial resistance poses a significant threat to the NHS>

AMR occurs when microorganisms (such as bacteria, viruses, fungi, and parasites) become resistant to antimicrobial medicines like antibiotics etc.
The World Health Organization (WHO) has declared AMR as one of the top 10 global public health threats.
Challenges include: longer hospital admissions, increased risk surgeries and higher death rates.
Healthcare strategies to tackle resistance: early prevention and timely diagnosis > relies on the work of the biochemists!

Drone delivery in the NHS >
In 2022, the NHS announced a ground-breaking trial where cancer patients would receive chemotherapy drugs via drones.
Drone delivery takes 30 mins compared to the usual 4 hours. This efficiency benefits patients and minimizes the risk of drug deterioration due to short shelf life.
The success of this trial could pave the way for same-day orders of vital medical treatments. This demonstrates the NHS’s commitment to ensuring timely treatment while also cutting costs and carbon emissions.
Drone technology could help England become a world leader in cancer care!

Epic Beaker IT System all over the NHS >
In 2023, Guy’s and St Thomas’ NHS Foundation Trust and King’s College Hospital NHS Foundation Trust jointly launched the Epic Beaker system.
This was the largest single go-live of Epic’s electronic patient record anywhere in the world.
It is one single, electronic IT system = It provides staff with a complete overview of a person’s care, freeing up more time for patient care.
Benefits: comprehensive data, efficiency and patient empowerment using MyChart app.
The successful implementation of Epic Beaker represents a significant step forward in healthcare technology, benefiting both patients and staff!

Question 45

Biomedical Scientist vs Clinical Scientist

Answer 45

Biomedical
-Highly trained in scientific methods in chosen discipline
- Mainly lab based, processing patient samples
- Performing diagnostic testing using manual methods or specialised equipment
- Their results provide basis of >75% of diagnoses

Clinical
-Evaluate, develop and implement diagnostic treatment methods
- Aiding the diagnosis and management of disease
- Often communicates directly with patients
- Advise on patient care working alongside clinicians
- Role varies between disciplines

Question 46

Who are IBMS?

Who are HCPC?

Answer 46

Institute of Biomedical Science

• Represents biomedical scientists (BMS) in the UK and internationally
• Support, progress and promote BMSs and the profession
  -Provides the educational and professional standards individual must meet to become a BMS

Health and care professions council

• Regulator of 16 health and care professions in the UK
• Sets standards of proficiency
• Approves training programmes
• Maintains register of individuals qualified to work

Question 47

Quality and Good Manufacturing Practice Training - Learnt at Fleet

Answer 47

ISO QUALITY STANDARDS
ISO9001 (general) and ISO13485 (medical devices such as in vitro diagnostics IVDs > hospitals). There is an overlap between the two.
Fleet examples: HIV and Troponin Ab conjugates for use in blood test assays.
Validating accuracy of DNaseB reagents on the Optilite machine for use in hospital labs to diagnose patients with streptococcal infection.

GOOD MANUFACTURING PRACTICE - GMPS
GMP = production standards required for regulated products e.g. things that go into blood tests.
Good GMP examples: qualified equipment, trained staff, controlled environment, following MBRs!
Fleet do pre-clinical work only > products do NOT directly end up in peoples bodies e.g. assays that support clinical work.

QUALITY FOR STAFF
Quality management systems (QMS) applies to everyone.

NON CONFORMANCES NCNS
Important to learn from them to not repeat the same mistake again.
Biggest NCN at Fleet between the period of Jan 2023 - May 2024 was 8 equipment failures, 4 of which were from wireless dataloggers.
Avoidable NCNs included: product labelling errors, losses of product due to AKTA fraction collector malfunctions e.g. a batch of Biotin:BSA loss 50% of fractions. These NCNs could have been avoided as were caused by human error.

QMS BASIC PRINCIPLES - IMPORTANT FOR AUDITS (mention on CV and in interviews)
1. Controlled Documents - such as SOPs, MBRs. These are compliant with ISO quality standards. Must make sure you use the current versions of these documents.

2. Quality Records - document the output of all procedures e.g. paperwork, lab book write up, equipment calibration records, scanning buffer sheets.
3. Traceability - means you have documented enough detail within our quality records to show exactly how the procedure was followed e.g. recording lab book write ups.

How we do things = controlled documents.
What we did = quality records.

LAB BOOK WRITE UP REQUIREMENTS
Record what actually happens > needs to be a high enough level of detail so that someone else would be able to follow it.
Record every step.
Include date and reference job number at beginning of work.
Sign off at the end of work and include date completed.
Get completed write ups reviewed as soon as possible.
Update index at start of lab book.

List all raw materials including name, supplier, cat code, batch / lot number and expiry date.
Raw materials that are made in house should be given an in house reference and expiry date.
Receipt of raw materials: Lab Acceptance Record > must be kept up to date.
List all equipment used.
Sign, initial and date errors made, pasted in paperwork and any blank boxes that were not used.
Use the 24hr clock for date format.
Write neatly and legibly.

EQUIPMENT
If you are an equipment owner, you must be familiar with the current version of the SOP for that piece of equipment and must make sure it is accurate!
SOPs also specify any required qualification, calibration, performance checks, maintenance and cleaning.
Everything gets at least minimal cleaning (a wipe-down with IPA every 6 months).
Must record the calibration and cleaning after completion.

ENVIRONMENTAL MONITORING
Fleet monitor the bioburden in the labs > it must be kept at an acceptably low level. A spike in bioburden bacterial and fungal data is cured by an extra lab cleaning being issues on top of the monthly one.

Question 48

What automated analysers do they use in Clinical Biochem and what do they test for?
Key - LEARN FOR INTERVIEW

Answer 48

Cobas 8000 analyser - Tests:

• Ion-selective electrode (ISE)module is used to measure
  the concentration of electrolyte such as Na+ and K+ in the
  sample by measuring the electrical potential difference.
• General chemistry module (c702) is used for measuring a
  wide range of analytes such as glucose, creatinine (kidney) and C
  reactive protein (liver = infection / inflammation) (CRP). Concentration of the analytes is
  measured with the colour intensity of the reaction mixture.
  Creatinine level in the blood or urine shows how well the kidneys are working
• Immunoassay module (e602) measures the concentration
  of analytes such as vitamin B12, ferritin using immunoassay.

Sebia capillary 3 TERA analyser:

*This analyser is used for measuring
the amount of glycated haemoglobin
(HbA1c) in patients’ samples.

• HbA1c can reflect the glucose level in
  patient’s blood over the past 1-2
  months.
• The analyser measures the HbA1c
  concentration by the capillary
  electrophoresis system.
• The measurement of HbA1c aids the
  diagnosis and monitor of diabetes
  mellitus.