Bioprocessing

1. Bioprocessing PHSC 438 October 2006 Dr. Tim Bloom

2. Overview • History • Product examples • Using microbial cells • Using mammalian cells • Preview of the rest of the semester

3. Biotechnology • Using biological systems for production • Using living cells/organisms • Using cellular components • Convert raw materials into product • Does not include agriculture/ranching • Unless organism is genetically modified • Golden rice • Growth-enhanced salmon

4. Bioprocessing • Branch of biotechnology • Using genetically engineered organisms • Contain foreign DNA • Usually codes a protein used as a product • Insulin • Human growth hormone • Microbial cells, mammalian cells, plants, intact animals

5. History of Biotech • First products not commercial • Beer, wine, vinegar • Bread and cheese • Micro-organisms convert raw materials into more useful product • Empirical development with local microbes

6. More Recent • Use enzymes to economize production • Proteases for leather • Amylases for simple sugars • Industrial scale enzyme production • Required reliable sources of enzymes • Cheap source is microbes • Growth and harvest techniques developed

7. Industrial Use of Enzymes • Enzyme trapped in a column • Raw materials passed through • Products formed by interaction with trapped enzyme • Product captured in eluant from column

8. The Golden Age • After World War II • Development of antibiotics • Produced by bacteria and fungi • Development of larger scale techniques • Tanks for growing cells • Systems for moving sterile liquids around • Systems for harvesting products from cells

9. Culture Scale Affects…

10. Microbial Products • Small molecules • Vitamins • Amino acids • Antibiotics • Alcohols • Organic solvents • Proteins • Therapeutic proteins • Reagent enzymes

11. Non-microbial Cells • Need for vaccines against viruses • Started with chicken eggs • Moved to cultured monkey cells • Developed systems for large-scale growth • More difficult than growing microbes • Continued development for economy of scale

12. Labor Intensive Viral Production

13. Mammalian Cell Products • Proteins • Antibodies • Cytokines • Growth factors • Viral vaccines • Also more economicalif grown in tanks

14. Moving Forward • Current efforts focused on improvement • Developing new products • Creating useful new cell hosts • Optimizing efficient growth and harvest of cells • Improving purification technology • More convenient formulations for administration

15. Process Development Overview • Engineer cells to make product • Optimize growth conditions • Develop purification protocols • Establish formulation

16. Engineering Cells • Insert foreign DNA into host • Codes for protein (product) • Codes for enzyme (makes product) • Codes for regulator (changes product level) • Bottom line: cell host makes more of a product that unmodified cell can

17. Growth Requires… • Nutrients • Dissolved oxygen • Waste removal (or dilution) • Appropriate pH and temperature • Supplied by medium • Controlled by (semi-) automated systems

18. Cell Culture Growth Curve

19. Product Purification • Separation from other things in system • Host cells • Host cell components • Media components undesired in final product • Based on chemical/physical properties • Interactions with other molecules (chromatography) • Size (filtration) • Solubility (precipitation)

20. Formulation • Excipients • Dosage form/administration route • Protect product • Physical degradation • Chemical degradation

21. From the Beginning… • Cells can be grown as individuals • Bacteria and yeast • Animal cells • Plant cells • How?

22. Next Time… • Kinds of cells used in industry • Requirements for industrial scale cell growth • Systems used to keep cells happy