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Bioprocessing. Michael Ladisch Laboratory of Renewable Resources Engineering Agricultural and Biological Engineering Weldon School of Biomedical Engineering - PowerPoint PPT Presentation
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College of AgricultureCollege of Engineering
Bioprocessing
Michael Ladisch Laboratory of Renewable Resources
Engineering Agricultural and Biological Engineering
Weldon School of Biomedical Engineering
Purdue University
Outline
1. Bioprocessing Defined
2. Pharmaceutical Industry Trends
3. Unit Operationsa. Bioreactorsb. Recoveryc. Rapid Prototyping
4. Opportunities in Discovery, Learning, and Engagement
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)
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.
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)
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)
Pharmaceutical Industry Trends, 2005Sales: Global $550 billion
US $246 billion
China $ 8 billion
7 of 30 top pharmaceuticals from biotechnology
Biogenerics likely slow to be introducedC and E News, Dec, 2005
Most Major Products in 2005 were Drugs
C and E News, Dec, 2005
Sales to June 2005 ($ Billions)
Cholesterol & triglyceride reducers $ 31.6
Antiulcerants 26.3
Antidepressants 20.1
Antipsychotics 15.5
Erythropoietins 12.1
Calcium antagonists, plain 11.9
Antiepileptics 11.7
Antirheumatic nonsteroidals 11.4
Oral antidiabetics 10.4
All other antineoplastics 9.9
TOP 10 THERAPY CLASSES $160.9
Blockbusters86 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
Metabolic Engineering
Redirecting pathways in an organisms to obtain more product
Yeast
Bacteria
Mammalian Cells
Plants and Animals
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
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
PDMS
SiO2
Flat PDMS cover
SiO2, glass, or PDMS substrate
Glass fiber (~12 μm diameter)
Labeled avidin (green) and BSA (red) liquid mixture; t=0
Glass fiber
t= ~3 minutes
Glass substrateSiO2 substrate
(a) (b) (c)
Flow channel
Rapid-prototyping
Huang et al., 20031 nL/mm channel
TAKES LESS THAN 15 minutes
Dip Coating Fibers with Protein or Particles
Ultra-sonic cleaning in EtOH for 5 min
EtOH
Dip-coat in protein or particle slurry
Glass fiber coated with particles
Glass fiber
Dimethyl-amino microbeads (0.8 µm diameter)
Stationary Phase on Fiber
OH OH OH+N CH3CH3
H
Bare glass fiber
Glass fiber pre-coated with biotin-BSA
Biotin
Streptavidin microbeads (0.8 µm diameter)
+
Streptavidin
Glass Fiber Coated with Streptavidin Beads
Commercially available Micro / nano Particles
Type Surface Chemistry Size
(µm)
Surface charge
Silica Hydroxyl -OH 0.1-7.9 -
Polystyrene Hydroxyl -OH 0.7-7.9 -
Polystyrene Carboxyl -COOH 0.7-7.9 --
Polystyrene Sulfonate -SO30.7-7.9 ---
Polystyrene Amino -NH30.7-7.9 +
Polystyrene Dimethylamino -NH(CH3)20.7-7.9 ++
Data from Spherotec, Inc. Libertyville IL.
Type Surface pI Size
(µm)
Surface Affinity
Polystyrene Antibody (IgG) NA 0.7-7.9 Binds protein A, G
Polystyrene Avdin 10.0 0.7-7.9 Binds to biotin
Polystyrene Streptavidin 5.0 0.7-7.9 Binds to bitoin
Polystyrene Biotin NA 0.7-7.9 Binds to strept/avidin
Polystyrene Protein A 4.9 0.7-7.9 Binds to IgG antibody
Polystyrene Protein G 5.0 0.7-7.9 Binds to IgG antibody
Data from Spherotec, Inc. Libertyville IL.
Commercially available Micro / nano Particles
Microscale Chromatography
Glass fiber coated with dimethylamino microbeads
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
Glass fiber coated with biotinylated BSA
Glass substrate Glass substrate
PDMSPDMS
t= ~3 minutes t= ~5 minutes(a) (b)
Huang et al., 2003
A B
CentrifugationCentrifugation
Fermentation or cell culture
Lysis
Centrifugation
Micro-filtrationMicro-filtration
Hold TankHold Tank
Precipitation TankPrecipitation Tank
Membrane SeparationMembrane Separation
Buffer TankBuffer Tank
Hold TankHold Tank
ChromatographyChromatography
Product Tank
Elution Buffer Tanks
AdsorptionAdsorptionAdsorption
Chromatography
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