BIOREACTOR SYSTEM DESIGN - bpceng.combpceng.com/docs/BPCiir981b.pdf · BIOREACTOR DESIGN...
Transcript of BIOREACTOR SYSTEM DESIGN - bpceng.combpceng.com/docs/BPCiir981b.pdf · BIOREACTOR DESIGN...
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BIOREACTOR SYSTEM DESIGN
CELL CULTURE SYSTEM DESIGN & SCALE-UP.INDUSTRY TRENDS FOR IMPROVED RELIABILITY AND
PERFORMANCE EQUIVALENCE.
By: Ted DeLoggio
BIOPROCESS CONSULTANTS
The Institute for International Research
BIOREACTOR SYSTEMS
Philadelphia, April 20, 1998
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CURRENT DESIGN TRENDS
MAJOR TOPICS
• PROCESS DESIGN & SCALE-UP• BIOREACTOR• PIPING• BIOREACTOR SYSTEM
INTEGRATION• BIOPROCESS SYSTEMS
INTEGRATION
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PROCESS DESIGN & SCALE-UP
COMMON CELL CULTURE SCALE-UP PROBLEMS“OBSTACLE OR OPPORTUNITY”
• [CO2] TOXICITY
• HYDRODYNAMIC SHEAR STRESS
• INTERFACIAL SHEAR
• BLEND TIME
• OTR REQUIREMENTS
• METABOLIC EQUIVELENCE
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[CO2] TOXICITY
DESCRIPTION OF PROBLEM[CO2] sensitivity begins at 75-100 mmHg
Gas exchange occursas bubbles rise. Sizeand composition ofbubbles change.
BALANCE
•OTR•CTR
Gas exchange at surface.
Cells consume O2and produce CO2.CO2 reacts with water to form HCO3
-
CO2O2
CO2O2
HCO3-
+H+
CO2O2
CO2O2
[CO2]74 ppm
OUROTR
CO2O2
CTRCER
CO2O2
air overlay ~0.05 VVM
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[CO2] TOXICITYSCALE-UP IMPACT OF “OUR” ON [CO2]
025
5075
100
125
150
0 2000 4000 6000 8000 10000 12000
Bioreactor Volume, liters
[CO
2], m
mH
g
0.5 OUR 1.0 OUR 2.0 OUR (mmol/L/hr)
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[CO2] TOXICITY
Design and Operating Actions to Reduce the Riskof [CO2] Toxic Effects
• PREVENTIVE ACTIONS– USE DRILLED HOLE SPARGERS (>3000l)– MAXIMIZE P/V, REDUCE SUPPLIMENTAL O2– IMPELLER TYPE, CONSIDER (K/Np) RATIO ↓– CONSIDER STRIPPING SPARGER IF SINTERED
ELEMENTS ARE USED
• REMEDIAL ACTIONS– EVALUATE BUFFER SALT COMPOSITION– CONTROLLED CARBON SOURCE ADDITION– CONSIDER USE OF STRIPPING GAS– OPTIMIZE pH CONTROL LOOP TUNING
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MIXING SCALE-UP PROCEDURES IN BIOTECHNOLOGY
All Procedures Require Geometric Similarity in Scale Up
CRITERIA PROCEDURE PROCESS EXAMPLE
Equal Mass Transfer Coefficients
(P/V)2 ≅ 1 (P/V)1
Microbial FermentationCell Culture
Equal Physical Shear to particles
(σ)2 ≅ 1 (σ)1
Cell culture
Equal Blend time or Vessel Turnover
(N)2 ≅ 1 (N)1
Rapid Kinetic Reactions
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DESIGN SENSITIVITY PRESERVING VESSEL GEOMETRY IN SCALE -UP
Intrinsic Dependencies of Highlighted Parameters
- Tank geometry- Sparge Rate & Bubble Size- impeller geometry- baffles & internals
OTR = kLaSPG/H (C* - CL) + kLaSUR/H (Cov - CL)
kL - mass transfer coefficient
a - air-liquid interfacial area
kLaSPG = C(P/V)α(vs)β
P0 = Np ρ N3 di5
shear rate = K Ndi
Mass Transfer and Shear Functionality depend on Vessel Geometry
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MAX
MIXINGESTABLISH THE REQUIRED AGITATION FOR YOUR SYSTEM
TipSpeed(ft/min)
1000M
500
CC
0
Mixing Requirements
1. Hydrodynamic Shear
2. OTR
3. Cell Suspension
MIN
VFD
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HYDRODYNAMIC SHEAR
The shear v. mass transfer scale-up dilemma
Shear Rate response to Equal P/V scale-upP/V=50 W/m^3
20KL5000L
1500L
600L100L
30L
3L
0
5
10
15
20
25
30
0 20 40 60 80 100 120
Bioreactor Diameter (in)
Shea
r Rat
e (1
/s)
Impeller Zone Shear Rate
Bulk Fluid Shear Rate
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Scale-up issues in Cell Culture-Mixing
Once the Design Basis for Mixing is defined (e.g. equal P/V), the controlling shear damage equation must be determined
• Average Shear Rate ~ KN
• Maximum Shear Rate ~ K’ND
• Cumulative Shear Stress
• ISF
• Kolmogoroff Eddy Scale
• Energy Dissipation
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INTERFACIAL SHEARDESCRIPTION OF PROBLEM
du/dxBulk Flow
INTERFACIAL SHEAR PROTECTION
Agent Cells Killed Cells Killedper bubble # per batch %
None 1600 >100PEG 1000 >100PVA 450 >100Methocel 30 45F-68 5 8
SECONDARY EFFECTS OF F68• Reduces KL (minor)• Increases a (major)• Cell membrane interaction• Influences foam formation
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EVALUATION OF O2 TRANSFER REQUIREMENT
OTRMECH > OURMET
OUR = ( u /YX /O) X= (4.5 x 10-10 mmolO2/cell/hr)(6 x 109 cells/L)= 2.7 mmol/L/HR THE GOAL
OTR = KLa /H*(C * - CL)H = 101330 Pa/atm / 86000 Pa-L/mmolO2(C * - CL) = (Ci –Co )/LN[(Ci –CL )/(Co - CL )](C * - CL) = 0.250 atm @ 21% O2(C * - CL) = 0.800 atm @ 60% O2 (50/50:air/O2)
KLa = 2.7/1.17*0.250 = 9.2 1/h air spargeKLa = 2.7/1.17*0.800 = 3.0 1/h 50/50 air/O2 sparge
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CURRENT DESIGN TRENDS
MAJOR TOPICS
• PROCESS DESIGN & SCALE-UP• BIOREACTOR• PIPING• BIOREACTOR SYSTEM
INTEGRATION• BIOPROCESS SYSTEMS
INTEGRATION
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BIOREACTOR DESIGN METHODOLOGY
GOAL:
Reproduce the metabolic and physical environment achieved at the current scale of operation to produce product at the new scale with equivalent quality and quantity. To assure safety, consistency, robustness, and validatability of the process per the customers
established product & process requirements.
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PROCESS OBJECTIVES
• Define mixing, mass transfer, heat transfer needs
• Define execution “building blocks”
• Define component and line sizing
• Define reliability, safety, cost & compliance needs
• Detail the equipment arrangement
BIOREACTOR DESIGN METHODOLOGYDESIGN MILESTONES
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BIOREACTOR DESIGN METHODOLOGYDESIGN MILESTONES
INTEGRATED PROCESS OBJECTIVES:
• FDA regulations - product quality and consistency
• Containment - product and worker safety
• Validation execution and documentation
• Maintenance Access
• SIP/CIP protocols
• Controls integration
• Downstream process integration
• Flow of materials and personnel
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BIOREACTOR VESSEL DESIGN
• DESIGN FOR PROCESS, CIP & SIP
• PRESERVE GEOMETRIC SIMILARITY– HL/Dt = 1.5 (cell culture)– HL/Dt = 2.5 (Microbial)
• NOZZLES– ORIENT TOP ASSEMBLY FOR SPRAYBALL COVERAGE.– USE “SHOP” WELDED VALVE ASSEMBLIES.– SLOPE TO DRAIN, AVOID DEADLEGS.– USE FLUSH MOUNTED SAMPLE AND HARVEST VALVES.
• USE DIMPLED OR HALF-PIPE JACKET DESIGN– CONSIDER TWO SIDE WALL ZONES.– CONSIDER ONE BOTTOM ZONE
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Large Scale Bioreactor DesignExample
• Operating Range• 18,000L –22,000L (Vw)• 28,000L (Vt)
• Aspect Ratio• HT/DT = 1.75• L/ DT = 1.43
• HL/DT• At 18,000L: 1.1• At 22,000L: 1.4
• Di/DT• Hydrofoil - 0.45• Hydrofoil - 0.45
• 4 Baffles • Width – 0.1DT• Clearance – 0.01DT
• Drilled Hole Sparge Element• 4 mm / 2 mm
193 (HT) 157 (L)
110 DT
HT= Total vessel inside height (in)
L = tangent to tangent height (in)
HL = Liquid height (in)
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AGITATION CONSIDERATIONS
• USE BOTTOM MOUNTED DESIGN IF POSSIBLE
• DOUBLE MECHANICAL SEAL vs MAGNETIC– MAGNETIC OPTIONS IMPOSE SCALABILITY ISSUES.
SUITABLE FOR PROCESS VESSEL NOT BIOREACTOR– USE PRESSURIZED BARRIER FLUID– SELECT “BEST” SEAL NOT BIOREACTOR VENDOR
• PROVIDE “ALARM & RECOVERY METHOD FOR SEAL FAILURE
• USE DIRECT SPEED PICK-UP FOR ACCURACY
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AGITATION CONSIDERATIONS
• IMPELLER TYPE, DIAMETER, AND NUMBER
• VESSEL, AGITATION, AND SPARGE DECISIONS ARE INTERDEPENDANT.
• CONSIDER ACCESS FOR MAINTENANCE
• SHAFT DESIGN & DEFLECTION CALCULATIONS.– Design for the worst case scenario and future impeller needs
• GEAR BOX FOR LOW SPEED OPERATIONS
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BIOREACTOR SYSTEM INTEGRATION
• VESSEL ADDITIONS
• GAS MANAGEMENT AND CONTROL
• TEMPERATURE CONTROL
• EXHAUST
• AGITATION
• PROCESS CONTROL AND SENSORS
“BUILDING BLOCKS”
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CURRENT DESIGN TRENDS
MAJOR TOPICS
• PROCESS DESIGN & SCALE-UP• BIOREACTOR• PIPING• BIOREACTOR SYSTEM
INTEGRATION• BIOPROCESS SYSTEMS
INTEGRATION
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PROCESS PIPING
• SKID v. D&B v. GRAY SPACE OPTIONS• DESIGN AND SIZE FOR PROCESS, CIP & SIP• LOCATE CRITICAL COMPONENTS FOR
ACCESSIBILITY• REDUCE FIELD HAND WELDS• SLOPE AND DRAIN LOW POINTS• CONSIDER 3-D MODELING FOR INTERFERENCE• INTEGRATE DRAWINGS WITH “BOM” AND “PM”• CONSIDER 3rd PARTY FOR INSPECTION
– USE PHYSICAL WELD SAMPLES– AGREE ON SPECIFICATIONS AND INSPECTION
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PROCESS PIPINGCIP DESIGN CONSIDERATIONS
• DEVELOP CIP SCENARIOS ALONG WITH PROCESS
• USE PARALLEL FLOW PATHS
• CONSIDER CIP SUPPLY AND RETURN FLOW REQUIREMENTS FOR LINE SIZING
• DEFINE STRATEGY TO VERIFY SECONDARY CLEANING PATH
• MODULARIZE AUTOMATION TO OPTIMIZE CLEANING CYCLES
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PROCESS PIPINGSIP DESIGN CONSIDERATIONS
• USE AUTOMATIC TEMP. MEASUREMENT AND STER. VERIFICATION
• ALLOW 8-12” DISTANCE BETWEEN RTD AND TRAP
• GROUP RTDs FOR EASE OF CALIBRATION
• USE VACUUM STEP TO ELIMINATE AIR
• CONSIDER STEAM ADDITION AND VENT LOCATIONS
• ENSURE ADEQUATE SUPPLY AND VENTING