lecture_2-biochemical processing
TRANSCRIPT
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BIOCHEMCIALPROCESSING:
OVERVIEW
CHARLES L. COONEY
DOWNSTREAMPROCESSING COURSE
MIT Professional Institute
August 1-4, 2005
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THE CHALLENGE OF
DOWNSTREAM
It is difficult to efficientlyand economically recover a
high purity biochemicalproduct from a complexmixture of related andfunctional molecules,
impurities and contaminants
which have similar physicaland chemical properties.
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YOUR GOAL
IF YOU DONT KNOW WHEREYOU ARE GOING AND YOUDONT HAVE A MEANS OFMEASURING WHERE YOU
ARE THEN YOU WONT KNOWWHEN YOU ARRIVE
Ama Dablam22,275 ft
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THE DIVERSE BIOCHEMICALPROCESS INDUSTRY
PRODUCTS AND SERVICES FORMULTIPLE MARKETS
Food & Beverage
Health CareTherapeuticsDiagnosticsDevice
Specialty ChemicalCommodity ChemicalsWaste Treatment
MANUFACTURING BY MULTIPLESYNTHETIC & EXTRACTIVE
TECHNOLOGIESBiosynthetic - Microbial, Animal, PlantExtractive Animal, PlantChemical Synthesis
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THE DIVERSE BIOCHEMICAL
PROCESS INDUSTRY (Continued)
PRODUCTS BELONG TO
MULTIPLE CLASSES Small Molecules Proteins
Nucleic Acids
Carbohydrates
Catabolites & Anabolites
Cells And Viruses
PRODUCTS & PROCESSESREGULATED FDA, EMEA
EPA
OSHA
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Where is the Leverage?Relationship of Profit to Price
PROFIT=
VF (SPSA
C )
Selling price isFixed by utilityAnd competition
Manufacturingcost
SpecificActivityof Product
Market share is afunction of proprietaryposition:
PatentsMarketingDistributionInnovation speed
Market size determinedby problem solved
Sa Sp Cm Profit(Units/lb) ($/unit) ($/lb) $ MM
200 0.45 55 210
200 0.45 35 330
200 0.35 55 90
500 0.45 55 1020
2000 0.45 55 5070
Sensitivity of Profit to Sp, Sp & Cm
V= 30 b lb sugar & Fm = 0.2
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POINTS TO CONSIDER IN
DOWNSTREAMPROCESSING
DSP begins with Raw Material selection Garbage inmeans garbage out
There are trade offs, e.g. between purity and yield NoFree lunch
Mass and Energy are conserved, What goes in mustcome out somewhere and in some form. There may betransformation in form
There are Impurities and Contaminants You will be watched, e.g. by customers (internal and
external) and the FDA. Therefore be sure to definemetrics and appropriate analytical methods
Regulation includes FDA, EPA, and OSHA
Design: Target the Spec Sheet Path the PFD Measure Analytical
Murphys Law Contaminants need control Lost material need robustness
PurityYield
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SUPPLY CHAIN IN
BIOPROCESSING
))()(cov(Re
))((
tivitySpecficAciTiterncyeryEfficie
eMarketSharMarket=
ManufacturingPlant Size
Utilities
Wastes
Market
Determines Plant Size
& Product Specs
Raw
MaterialsSynthesis DSP
Final
Product
Figure by MIT OCW.
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MANUFACTURING
BY FERMENTATION
Product
Raw Materials
Air
Steam
Water
Inoculum
Process Aids
Waste
Heat
Fermentor
Downstream
Processing
Process
Control
Figure by MIT OCW.
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WHEN SELECTING UNITOPERATIONS THERE ARECHOICES AND DECISIONS
MUST BE MADE
Cell Harvesting
Bioreactor
1. Centrifugation
2. Microfiltration
3. Ultrafiltration
Intracellular
Products
Extracellular
Products
Biomass Removal
1. Vacuum Filtration
2. Centrifugation
3. Microfiltration
4. Ultrafiltration
5. Press Filtration
6. Candle Filtration
7. Flotation
Cell Disruption
1. Homogenization
2. Bead Milling
3. Osmotic Shock
Cell Debris Removal
1. Centrifugation
2. Microfiltration
3. Vacuum Filtration4. Press Filtration
Renaturation
1. Solubilization
2. Reoxidation
Dehydration or Solvent Removal
1. Spray Drying
2. Freeze Drying
3. Fluid Bed Drying
Product Extraction By
1. Organic Solvents
2. Polymer/Polymer
3. Polymer/Salt
4. Supercritical Fluids
5. Adsorption
6. Reverse Micelles
7. Distillation
Concentration
1. Ultrafiltration
2. Evaporation
3. Reverse Osmosis
4. Precipitation
5. Crystallization
6. Extraction
6. Adsorption7. Distillation
Final Purification
Requirements
Contemporary Downstream Processing
of Biological Materials
Denatured Products
Requirements
Low PurityHigh Purity
1. Adsorption
2. Gel Filtration
3. Diafiltration
4. Electrodialysis
5. Electrophoresis
Figure by MIT OCW.
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Fermentation ProcessDevelopment
Metabolic pathwaysrocess kinetics
Elemental balances
Solubility & Equilibria
Mass transferWater balanceEquipment correlations
A +B --> C
Set of Enzymes and
Reactions
A + C ---> D +E
Net Biochemistry
CellEnvironment
Glucose
CO2
NH4+
O2
Product
H2O
Reactor
+ Utilities
Flow
pH
D.O.
Pres.
Air
Wt
Power
Comp.
Air
Comp.
D.O.
pH
Pres.
Flow
CornSryup Water
Flow
Wt
Wt
Product
CoolingSystem
Flow
NH3
Flow
Flow
Flow
Anti-foam
Temp
Temp
Temp
Performance Assessment
Growth Rate
Product Concentration
By-Product Concentration
Raw Materials Utilization
Experimental Parameter
Host cell selectionExpression system
Media design
Fermentation conditions
Aeration strategy
Cell harvesting strategy
Molecular Biology
Expression system
Plasmid design and copy number
Control of metabolism
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STRATEGIES FOR
MEDIA DESIGN Selection of media from
literature
Analogy with medium foranother organism
Rationale design from cell
and product needs andprocess demands
Experimental design
Who should be involved in media design?MicrobiologistPurchasingAnalytical chemist
Process engineering
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MEDIA DESIGN
MEETING THE REQUIREMENTS FOR GROWTH
PRODUCT FORMATION
(continued)
4. REGULATORY CONSTRAINTS
Qualification of vendors
Multiple sources
Traceability
Potential impurities or contaminants
Consistency
5. TECHNO-ECONOMIC CONSTRAINTS
Cost
Materials availability
Product recoveryEnvironmental impact
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FERMENTATION
MEDIANUTRIENT RAW MATERIAL PRETREATMENTCARBON SOURCE
GLUCOSE CERELOSE HYDOLYZED FROM STARCH
MOLASSES INVERSION
(SUCROSE TO FRUCTOSE AND GLUCOSE)
STARCH SOLUBILZIATION
CELLULOSE GRINDING AND HYDROLYSIS
FATS/OILS SOYBEAN OIL
COTTONSEED OIL
NITROGEN SOURCE
AMMONIA
PROTEIN HYDROLYSATES ACID OR ENZYME CATALYZED
HYDROLYSIS
COST
PURITY
CORN SUGAR
STARCH
MOLASSES
CELLULOSIC BIOMASS
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OVERVIEW OF MEDIADESIGN
There is a criticalneed for analyticalsupport
Raw MaterialOptions
FermentationProcess
DownstreamProcess
ProductSpecifications
Waste
TreatmentCost of raw materialCost of fermentation efficiencyCost of downstream efficiency
Cost of waste treatmentCustomer demands for product quality
5Cs
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FERMENTATION MEDIUM
COMPONENTSPENICILLIN
Molasses0.2% Soybean Oil
1% Cottonseed Flour
STREPTOMYCIN2.5% Cerelose4% Soybean Oil
LACTIC ACID BACTERIAPhosphate buffer0.5% Tryptone0.5% Yeast Extract
BACITRACIN3% Corn Steep3% Glucose
Bakers YeastMolasses
HighImpuri
ty
Content
Multiphase
Compone
nts
Variable
Quality
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