Biotechnology

Question 1

Define biologics

Answer 1

Any virus, therapeutic serum, toxin, antitoxin, used in the prevention, treatment or cure of diseases in humans

Question 2

How are small molecules developed?

Answer 2

Through high throughput SCREENING.

Question 3

How are biologics developed? Steps?

Answer 3

CONSTRUCT instead of screen
1.choose a target
2. Construct biologic candidates
3. Experimental testing
4. Clinical trials

Development, Validation, Manufacture, Formulation

Question 4

When is Q5D and Q5E quality control used

Answer 4

Q5D
- used when deriving new substrates for biotechnology purposes

Q5E
- used when making biosimilars

Question 5

What are the 3 enzymatic regulatory activity?

Answer 5

1. Replacement of deficient/abnormal proteins
2. Augment existing pathways
3. Novel function or activity

Question 6

enzymatic regulatory activity:
What do we use replacement of deficient/abnormal proteins for? (3)

Answer 6

a. hormone deficiencies:
- insulin, genotropic for (hGH deficiencies)

b. Hemostasis, thrombosis;
- clotting factors (8, 7a)

c. Gi disorders
- lactaid
- pancreatic enzymes

Question 7

enzymatic regulatory activity:
How do we augment existing pathways?

Answer 7

a. Anemia
- augment the erythropoiesis pathway

b. Immunoregulation:
- produce WBCs, platelets, and modulate immune response

Question 8

enzymatic regulatory activity
How do we use novel function/activity?

Answer 8

a. Degrade existing molecules
- disrupt neuromuscular junction, lung mucus viscosity reduction

b. hemostasis, thrombosis
- produce anticoagulants

Question 9

In special targeting activity:
How do we interfere with molecule/organism

Answer 9

a. Prevent infectious diseases
- Stop HIV entry

B. Cancer
- bind surface receptors of cancer cells

c. Immunoregulation
- Bind TNF, prevent inflammatory cascade

Question 10

In special targeting activity:
How do we deliver molecules

Answer 10

Cancer
- deliver cytotoxic agents to cancer cells

Question 11

What type of cells produce antibodies?

Answer 11

B cells

Question 12

What is the structure of antibodies? Fc vs Fab

Answer 12

Large Y shape
4 protein chains connected by disulfide bonds

Fc (crystallisable) = constant region
Fab (antigen binding) = variable region

Question 13

How do hypervariable regions in FAB work?

Answer 13

Target specific parts of antigens in a “lock and key”

Question 14

Define polyclonal antibodies

Answer 14

generated from an Immune response to an antigen
- Different B cell receptors produce a range of antibodies
- multiple lineages of B cells = polyclonal antibodies

Question 15

Define monoclonal antibodies

Answer 15

single line of B cells, selected to produce the SAME antibodies (-mab)

Question 16

In humanization of antibodies define
Chimeric
Humanized
Human

Answer 16

FC region from mice leads to shorter half life, less immune response, inefficient effect

• Chimeric: Human FC + mouse-derived FAB (variable) region -OMAB
  
  • Bad for half life, immune response
• Humanized: human Ab + mouse CDR -IMAB
• Human: no mouse genetic sequences -ZUMAB

Question 17

Define antibody drug conjugates
Characterisitics?

Answer 17

Small molecule is attached to mAB to make use of its specificity
- can be cleavable, non-cleavable
- Attachment site can be Fc or FAB

Question 18

What do the following excipients do to aid stability/delivery

Salts
Surfactants
Buffers
Cyclodesxtrins
Polyalcohols
PEG

Answer 18

Salts
- decrease denaturation, aggregation

Surfactants
- prevent adsorption of proteins

Buffers
- maintenance of pH

Cyclodesxtrins
- decrease aggregation

Polyalcohols
- stabilize, maintain isotonicity

PEG: INCREASE HALF LIFE OF PROTEINS DRUGS

Question 19

Describe lyophilization. Issue?

Answer 19

“freeze drying”
- remove water from product –> form dry powder. Good for long-term storage

Issue: protein aggregation
- stabilizers (sucrose)
- Surfactants
- DO NOT SHAKE

Question 20

What is Q5C
Define potency and purity in this

Answer 20

Q5C
- stability testing of biotechnological/biological products

Potency: ability of a product to achieve its intended effect
Purity: relative, dependant on the product

Question 21

Which protein takes the longest to develop in vaccines? quickest?

Answer 21

Longest: protein VLP
quickest: mRNA

Question 22

Define antisense gene therapy

Answer 22

Antisense RNA binds to sense sequence to block protein synthesis

Question 23

Define cell therapy

Vivo cell therapy?
Allogenic cell therapy?

Answer 23

Using THE WHOLE CELL to bring effect
* Blood transfusions, bone marrow transplants, stem cells for regenerative medicine

Ex vivo cell therapy: modifying the patients OWN cells but OUTSIDE the body

Allogenic cell therapy: using cells from a DONOR

Question 24

Differentiate between somatic cells and germline cells in genome editing

Answer 24

Changes to somatic cells are not passed on to offspring,
Changes to germline cells are passed to offspring

Question 25

What does CRISPR stand for? Cas9?

Answer 25

CRISPR = Clustered regularly interspaced short palindromic repeats

Crispr associated system