Microbial Insulin Production powerpoint presentation

Transcript

1. None

2. contents • Introduction • History • Structure, function and synthesis

   of insulin • Insulin production by Recombinant DNA technology: Two-chain method • Bio-production Steps • Different Expression Systems of insulin • Conclusion • References 2

3. Introduction rapid increase in the number of diabetic patients limitation

   in production capacity high production cost the demand for recombinant insulin Recombinant human insulin has been produced predominantly using E. coli and Saccharomyces cerevisiae for therapeutic use in human. 3

4. History 4

5. Biopharmaceutical products • Therapeutic monoclonal antibodies • the hormones •

   growth factors 5

6. Structure and function of insulin 6

7. Insulin synthesis 7

8. Insulin production by Recombinant DNA technology: Two-chain method 8

9. Bio-production Steps 1. Gene Isolation 2. Insertion into the Plasmid

   9

10. Bio-production Steps 3. Transfection 4. Medium Preparation 5. Bioreactor Fermentation

    10

11. Bio-production Steps 6. Crude Product Isolation 7. Purification 9. Chain

    Joining 11

12. Different Expression Systems of insulin • E. coli • Yeast:

    • Saccharomyces cerevisiae • Pichia pastoris • Transgenic Plants: • Arabidopsis thaliana • Tobacco and Lettuce Plants • Strawberry Plant 12

13. E. coli • the first expression system used to produce

    human insulin in 1978 • Advantages • Disadvantages • new technologies to solve problems 13

14. Yeast • another commercially-used expression system for heterogeneous protein production

    with recombinant technology because of its PTMs. • Advantages • Disadvantage: proteins synthesized in yeast have high-mannose N-glycosylation activates an immune response in humans shortens the half-life of recombinant proteins in vivo 14

15. Yeast: Saccharomyces cerevisiae • Produce: • Proinsulin • Fast-acting insulin

    • Long-acting insulin 15

16. Yeast: Pichia pastoris • presence of the methanol-inducible alcohol oxidase-1

    promoter (AOX-1) • Heterogeneous protein expression attained by P. pastoris is approximately 30% of its total cell protein 16

17. Transgenic Plants • Plants do not have human pathogens and

    because they are eukaryotic, their PTM machinery is more similar to humans than that previously described in yeast. • Arabidopsis thaliana • Tobacco and Lettuce Plants • Strawberry Plant 17

18. Conclusion Dietary and lifestyle changes are causing dramatic increase in

    diabetes incidence all over the world. Both Type I and Type II diabetic patients use insulin, however late stage Type II diabetes patients require large doses of insulin as they develop insulin resistance. Current manufacturing technologies will not be able to meet the growing demand of insulin due to limitation in production capacity and high production cost. 18

19. References Baeshen, N. A., Baeshen, M. N., Sheikh, A., Bora,

    R. S., Ahmed, M. M. M., Ramadan, H. A., ... & Redwan, E. M. (2014). Cell factories for insulin production. Microbial cell factories, 13(1), 1-9. Alyas, J., Rafiq, A., Amir, H., Khan, S. U., Sultana, T., Ali, A., ... & Ahmad, A. (2021). Human Insulin: History, Recent Advances, and Expression Systems for Mass Production. Biomedical Research and Therapy, 8(9), 4540-4561. Riggs, A. D. (2021). Making, Cloning, and the Expression of Human Insulin Genes in Bacteria: The Path to Humulin. Endocrine Reviews, 42(3), 374-380. 19