Biomanufacturing Jim DeKloe

1. Biomanufacturing Jim DeKloe

2. Biotechnology is the intersection of • Science • Business • Government

4. Traditional Pharmaceutical “Big Pharma” Drugs Small molecules Produced by Chemistry Example: Aspirin Biotechnology companies “Biotech” Proteins Large Molecules Produced by Living Organisms Example: EPO Drugs versus “biotech”

5. Drugs Small molecules Produced by Chemistry Simple Pure Molecules Easy to Test Quality Example: Aspirin Biologics Complicated Mixture of Large Molecules Difficult to Test Purity Emphasis on Consistent Process Examples: Vaccines, Plasma and Serum History

6. Technician Training Biotech clusters thrive when all elements of the industry are represented Until recently, the emphasis on R & D meant that formal training targeting technicians was rare Community Colleges have moved into this niche Technician training will be even more important as more companies move from R & D into biomanufacturing

7. A Model Academic - Industry Collaboration • Building biomanufacturing curriculum • Tailoring curriculum to develop specific required skills • Incorporating regulatory requirements into curriculum

8. Genentech supplied • Sabbatical Experience • Expertise • Donated Equipment • Presence on Advisory Committee • Guest Speakers

9. Traditional versus Industrial Traditional Industrial Education Training Lab Atmosphere Pilot Plant Atmosphere Set Times Shifts Lab Protocols SOP Lab Notebook Batch Records Individual Team

10. Question Authority! • Change Academic Paradigm • Borrow vocational tech and business ideas • Teach skills as well as educate • Emulate the industrial experience • Share the ideas that work

11. Back to Basics - Cells

12. Eukaryotic Cells can be Engineered Also

13. Protein! • The Protein is the Product!

14. Protein Structure • Proteins can be modified after they are synthesized • The addition of sugars – glycosylation – may be important – it affects solubility and pharmacokinetics (blood half live) • Glycosylation patterns may vary • Repeat because it is so important: If a protein loses this shape, it loses its function

15. Protein! • The protein must not be unfolded (denatured) or it loses its function. Proteins can be denatured by: • pH extremes • temperature extremes • organic chemicals • agitation

16. Protein! • DNA is the Flash • Protein is the Cash

17. Protein! • DNA is the Show • Protein is the Dough

18. Protein! • DNA is the Bling • Protein is the Thing

19. Bacterial Cells – Prokaryotic Cells

20. Cells are grown in a large scale

21. Upstream • Cells Divide and are transferred to larger and larger volumes • Cells are induced to produce protein • Many in-process samples are taken; some are sent to QC for a variety of tests. For mammalian cells, they will check for contamination by viruses and mycoplasma

22. Recovery - Downstream • Cells are separated from the medium surrounding them by filtration or centrifugation. • Typically: • In E. coli culture (slang is fermentation), the cells are retained and the medium is discarded • In cell culture (CHO cells), the cells have been engineered to secrete the protein, so the cells are discarded and the protein-containing medium is retained

23. Protein Purification • The protein of interest must be separated from the other proteins in the cells or medium until it is 99.99% pure. • This is accomplished by ultrafiltration and chromatography

24. Ultrafiltration • In ultrafiltration the protein containing solution is passed through a filter with pores of a defined size. This can separate proteins of different sizes from one another, or can separate protein from medium components.

25. Chromatography • Chromatography methods use resins of different types to exploit the characteristics of the protein to interest to separate it from other contaminating protein

26. Types of Chromatography • Type of Chromatography Protein Property • Gel Filtration (Size Exclusion) - Size • Ion Exchange (Cation or Anion) – Charge • Hydrophobic Interaction Chrom. – Hydrophobicity • Affinity Chromatography - Function or Special Characteristic

27. Formulation • Once the protein has been purified, the bulk purified protein will be placed in the formulation buffer and sterile filtered. During formulation, adjuncts are added to: • Stablize pH • Adjust osmolarity • Prevent aggregation • Cryoprotect (for lyophilized products)

28. Fill • Fill is completed in a special facility that has highly filtered air, gowned personnel with special training, This process is highly automated. • A measured amount of the formulated protein is aliquoted into sterile ampules

29. Finish • The protein may be lyophilized (freeze dried). This makes it more stable, but physicians may find this form of the drug less convenient to use. • Closures are added • Labels are applied.

30. Quality Assurance • The law requires a QA/QC department that is separate from manufacturing • They will test: • Raw Materials • In-Process Samples • Utilities • Environmental Monitoring • The final protein product

31. Regulation • In the United States, the Food and Drug Administration regulates pharmaceutical production • Part of the executive branch • Regulates $ 1 trillion of commerce a year • Center for Drug Evaluation and Research • Center for Biologics Evaluation and Research

32. What Are GMPs?