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Explore the intersection of science, business, and government in biotechnology. Learn about biomanufacturing of proteins and small molecules, emphasizing consistency and quality testing. Discover the importance of technician training and academic-industry collaborations to tailor curriculum for specific skills. Understand the processes of cell growth, protein purification, and formulation in biomanufacturing.
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Biomanufacturing Jim DeKloe
Biotechnology is the intersection of • Science • Business • Government
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”
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
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
A Model Academic - Industry Collaboration • Building biomanufacturing curriculum • Tailoring curriculum to develop specific required skills • Incorporating regulatory requirements into curriculum
Genentech supplied • Sabbatical Experience • Expertise • Donated Equipment • Presence on Advisory Committee • Guest Speakers
Traditional versus Industrial Traditional Industrial Education Training Lab Atmosphere Pilot Plant Atmosphere Set Times Shifts Lab Protocols SOP Lab Notebook Batch Records Individual Team
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
Protein! • The Protein is the Product!
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
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
Protein! • DNA is the Flash • Protein is the Cash
Protein! • DNA is the Show • Protein is the Dough
Protein! • DNA is the Bling • Protein is the Thing
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
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
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
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.
Chromatography • Chromatography methods use resins of different types to exploit the characteristics of the protein to interest to separate it from other contaminating protein
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
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)
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
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.
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
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