lecture_2-BIOCHEMICAL PROCESSING

8/7/2019 lecture_2-BIOCHEMICAL PROCESSING

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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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lecture_2-BIOCHEMICAL PROCESSING

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