Biotechnology in Use

Question 1

Summarise the examples covered in previous lectures of proteins and what they are best produced in

Answer 1

Insulin
- No post-translational modifications
- Best produced in bacteria/prokaryote

EPO
- Glycosylation (post translational modification)
- Best produced in mammalian cell culture/CHO/eukaryote system

Antithrombin
- Carboxylation (post translational modification)
- Best produced in transgenic animal/goats/pharming

Question 2

How can we design new proteins with new and/or improved function?

Answer 2

By protein engineering

Why?
- Improved therapeutics
- Industrial application
- Biotechnology and Biomedical research
- Drug discovery and development
- Agricultural and environmental applications
- Precision medicine
- Biodefence and Biosecurity

Question 3

How can we design new proteins with new and/or improved function?

Answer 3

Protein engineering:
1. Identify a need
2. Target selection (modifying an existing protein or making a new one?)
3. Design the protein
4. Gene synthesis
5. Expression and purification
6. Characterisation and optimisation
7. Validation and testing

Question 4

What are some examples of proteins used in biotechnology that answer some prominent questions?

Answer 4

How can we provide a permanent treatment?
- Use patients as the host for protein production
- Gene therapy for diabetes?

How can we continue to feed a growing population?
- Next generation dairy products
- Climate-friendly plants that make dairy proteins

Question 5

Describe gene therapy

Answer 5

Using viral vectors to insert recombinant gene into patient cells
- Uses viruses that only have the ability to bind to the cell surface and inject DNA (but in this case the only DNA in there is the viral vector)

Two types of viral vector:
- Integrating: integrates into host cell vector
- Non-integrating: injected into host cells but stays outside of genome and is still expressed

This process means if a person doesn’t have a functional gene for a particular protein, you can give it to them (permanent)

Question 6

What is the solution that gene therapy provides for Type I diabetes?

Answer 6

Problem: Pancreatic beta-cells destroyed
- antibodies recognise proteins on beta cell surface
- no insulin produced

Solution: reprogram pancreatic alpha-cells to behave as beta-cells

Question 7

Describe next generation dairy molecules

Answer 7

Casein milk powder currently produced via the dairy industry (approx. 80% of milk protein is casein)
- Want to madd produce this protein instead of using cows because they are bad for the environment and expensive

Miruku solves many problems currently surrounding milk and the environment.
- Uses transgenic plants to make the casein protein. The gene is expressed in the seeds so these are harvested and the protein is isolated from them.
- Solves climate problems and increases ability to feed growing population
- Proof of concept was taken from pork myoglobin protein in soybean