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Bioprocessing

Bioprocessing. Michael Ladisch Laboratory of Renewable Resources Engineering Agricultural and Biological Engineering Weldon School of Biomedical Engineering

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Bioprocessing

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  1. Bioprocessing Michael LadischLaboratory of Renewable Resources Engineering Agricultural and Biological Engineering Weldon School of Biomedical Engineering Purdue University

  2. Outline • Bioprocessing Defined • Pharmaceutical Industry Trends • Unit Operations a. Bioreactors b. Recovery c. Rapid Prototyping • Opportunities in Discovery, Learning, and Engagement

  3. Biotechnology Broadly defined in 1991 any technique that uses living organisms (or parts of organisms) to: 1. make or modify products, 2. improve plants or animals, 3. develop microorganisms for specific uses (Office of Technology Assessment, 1991)

  4. New Biotechnology Technology for manipulating genetic information and manufacturing products that are of biological origin or which impact biological activity. Based on methods introduced since 1970, applied in the laboratory since 1973, used on an industrial scale since 1979.

  5. Biotechnology Broadly defined in 2001 Technology with one or both characteristics: 1. uses organisms, or tissues, cells, or molecular components derived from living things, to act on living things 2. acts by intervening in the workings of cells or the molecular components of cells, including their genetic material. (National Research Council, Opportunities in Biotechnology for Future Army Applications, 2001)

  6. Bioprocess Engineering Subdiscipline within biotechnology responsible for translating life-science discoveries into practical products, processes, or systems capable of serving the needs of society (National Research Council, Putting Biotechnology to Work, 1991)

  7. Pharmaceutical Industry Trends, 2005 Sales: Global $550 billion US $246 billion China $ 8 billion 7 of 30 top pharmaceuticals from biotechnology Biogenerics likely slow to be introduced C and E News, Dec, 2005

  8. Most Major Products in 2005 were Drugs C and E News, Dec, 2005

  9. Blockbusters 86 products with sales in excess of $ 1 billion Potential for huge sales: smoking cessation inhaled insulin cervical cancer/human papilloma virus vaccines hypertension Niche blockbusters for cancer treatment (many monoclonal antibodies) C and E News, Dec, 2005

  10. Metabolic Engineering Redirecting pathways in an organisms to obtain more product Yeast Bacteria Mammalian Cells Plants and Animals

  11. Bioprocess Unit Operations: Bioreactors Microbial fermentation vs cell culture Recovery of Products Centrifugation Protein A (Affinity Chromatography) Ultra-filtration / micro-filtration Ion exchange, Size Exclusion Chromatography

  12. Bioprocessing: Cell Culture Important Produces monoclonal antibodies (MW 150 kD) Bioreactor size of 200 to 10000 L Media composition, avoiding contamination, supplying oxygen, removing wastes, operating bioreactor are challenges

  13. Rapid-prototyping Flat PDMS cover Glass fiber (~12 μm diameter) SiO2, glass, or PDMS substrate Labeled avidin (green) and BSA (red) liquid mixture; t=0 SiO2 substrate Glass substrate (c) (b) (a) t= ~3 minutes PDMS Flow channel SiO2 Glass fiber 1 nL/mm channel Huang et al., 2003 TAKES LESS THAN 15 minutes

  14. Dip Coating Fibers with Protein or Particles Glass fiber EtOH Ultra-sonic cleaning in EtOH for 5 min Dip-coat in protein or particle slurry Glass fiber coated with particles

  15. Stationary Phase on Fiber Dimethyl-amino microbeads (0.8 µm diameter) + OH OH OH H N Bare glass fiber CH3 CH3

  16. Glass Fiber Coated with Streptavidin Beads Streptavidin microbeads (0.8 µm diameter) Biotin + Glass fiber pre-coated with biotin-BSA Streptavidin

  17. Commercially available Micro / nano Particles Data from Spherotec, Inc. Libertyville IL.

  18. Commercially available Micro / nano Particles Data from Spherotec, Inc. Libertyville IL.

  19. Microscale Chromatography Labeled avidin (green; 10 µg/ml) and BSA (red; 10 µg/ml) liquid mixture; t=0 Labeled avidin (green; 10 µg/ml) and BSA (red; 10 µg/ml) liquid mixture; t=0 PDMS PDMS Glass fiber coated with biotinylated BSA Glass fiber coated with dimethylamino microbeads Glass substrate Glass substrate (a) (b) t= ~3 minutes t= ~5 minutes Huang et al., 2003

  20. Fermentation or cell culture Lysis Centrifugation Adsorption Adsorption Centrifugation Micro-filtration Elution Buffer Tanks A B Hold Tank Precipitation Tank Membrane Separation Chromatography Chromatography Buffer Tank Hold Tank Product Tank

  21. Bioprocessing Opportunities Biology component needed for designing batch unit operations Rapid Prototyping (microscale) will enable separations development Biorecovery and bioseparations engineering important Learning at P -12 starts with manufacturing cures

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