purification scale-up and technology transfer for lonza...

Purification Scale-Up and Technology Transfer for Lonza Biologics 20,000 L cGMP Facility

IBC Bioprocess International Conference & Exhibition 21st September, 2005

David PageLONZA Biologics, Portsmouth, NH

Facility Background

20,000 L LSBO manufacturing facilityMulti-product facility3 production bioreactors

Design CapabilityManufacture of antibodies and other recombinant proteinsDesigned to accommodate 2 g/L fermentation titresBatch harvested every 5-6 daysDesigned to process 4 chromatography stepsDesigned to process 3 UF stepsVI hold and VRF Bulk Fill operation

20,000 L Schematic

Centrifuge

FinishedGoods

Distributed toCustomers

FinalFiltration

2m column

Media Prep

Viralfiltration

FEEDS FEEDSFEEDS

Inoculum

Depth Filtration

Buffer Prep/Hold

0.22umFiltration

QC/QA

HarvestTank

2m column

1.4m column

1.4m column

1.4m column

TRAIN A TRAIN B TRAIN C

Conc/Diafil

Conc/Diafil

SUITE D SUITE C

SUITE CSUITE BSUITE AConc/Diafil

Conc/Diafil

Downstream Facility Photos

Suite B 1.4 & 2.0m columns Suite B UF skids

Downstream Overview

Chrom 1

Purification HoldTanks

SUITE A

Chrom 3

UF Concentration/Diafiltration

VRF

SUITE C

Chrom 4


Bulk Fill

SUITE D

Chrom 2


SUITE B

Tech Transfer Overview

• Key activities:1. Gap Analysis2. Pilot evaluation3. Process Design4. Facility Start-Up5. Development Runs/Scale-Up

– Training– Data Comparability

6. cGMP Engineering Runs– Process Description – Batch Records

7. Process Validation8. Commercial Production

Gap Analysis

Identify critical operations that impact facility fiti. Collection & Buffer Tank Constraintsii. Filtration area requirements (VRF, membrane filters)iii. Manual vs. Automated Controls Systems (Delta V

capabilities)iv. Collection Criteria (UV sensor path length)v. Column Packing Requirements & Methodsvi. Additional Equipment Required

PilotEvaluation

ProcessDesign

GapAnalysis

FacilityStart-Up Consistency CommercialDev.

RunsEng

Runs

Pilot Evaluations

• Aims & Objectives– Gain operations experience and product characteristics– Obtain sizing data for filters– Evaluate new/modified unit operations

• Equipment was a direct scale-down of LSBO equipmenti. 40cm Euroflow Resolute columnii. Euroflow packing skid and hydraulics

– Reproducible methods required for large scale– First time success is critical (product changeover)

iii. 0.5m2 flat sheet UF membranes

PilotEvaluation

ProcessDesign

GapAnalysis


RunsEng

Runs

Pilot Evaluations (cont.)

• Obtained filter sizing data for key steps• High throughput 0.2µm filters required for scale-up

– Optimization required for facility fit – Filter 1 selected to minimize filtration surface area

Capture Column Guard Filtration

0.0

10.0

20.0

30.0

40.0

50.0

60.0

Filter 1 Filter 2 Filter 3 Filter 4

No.

30"

Ele

men

ts

6 x 30” housing avail.

PilotEvaluation

ProcessDesign

GapAnalysis


RunsEng

Runs

Pilot Evaluations (cont.)

• UF optimization– New unit operation for required facility fit– Operate within membrane controlled (linear) region– Confirm flux decay to determine processing time

0.0 0.5 1.0 1.5 2.0 2.50

50

100

150

200

250

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350

0

50

100

150

200

250

300

350

Per

mea

te F

lux

(L.m

-2.h

-1)

T ransm em brane Pressure (bar)

420 L.m -2.h-1 Crossflow 480 L.m -2.h-1 Crossflow

PilotEvaluation

ProcessDesign

GapAnalysis


RunsEng

Runs

Process Design• VRF filtration

– Design filtration scheme to fit the facilityi. Confined to a filter skidii. Disposable manifold for flexibility (multi-product)iii. Tank to tank flow pathiv. Uniform flow to each filter element

PilotEvaluation

ProcessDesign

GapAnalysis


RunsEng

Runs

Process Design (cont.)

• Custom manifold design– Both in-house designs required for contingency– Demonstrated that both are operationally viable– Modular, autoclaved, disposable and SIP’d onto filter skid

PilotEvaluation

ProcessDesign

GapAnalysis


RunsEng

Runs

Process Design (cont.)

• Considerations– Request auto-cad drawings– Approve designs 3 months prior to implementation

– Field verify all designs and flow paths– Careful attention to lead times

– Build in safety margin for design and raw materials– Build in contingency for stable hold points

PilotEvaluation

ProcessDesign

GapAnalysis


RunsEng

Runs

Start-Up Activities

• Column packing performed at scale 6 months prior– HETP test method optimized– Evaluate range of bed heights for each column

• Wet-test of equipment– Test/modify logic parameters– Performed for untested unit operations

i. VRF step (transfer parameters)ii. UF (new skid, fed-batch operation)

PilotEvaluation

ProcessDesign

GapAnalysis


RunsEng

Runs

Manufacturing Campaigns

• OverviewDevelopment runs

Processed with draft batch records (BR)cGMP Engineering runs

Manufacturing operations compliant with cGMPQA approved BR’s

Process Consistency

Development Runs & Scale-Up

• Aims and ObjectivesTest and optimize unit operationsPerform operations within defined parametersIdentify and implement modificationsDeliver a fixed process going into cGMP Engineering runsEstablish a process that is

characterizedrobust reproducible

PilotEvaluation

ProcessDesign

GapAnalysis


RunsEng

Runs

Development Runs & Scale-Up (cont.)

• Optimize operationsSkid priming (air removal)Determine back-pressure within process lines (fine tune pressure set-points)Maximize recoveries (flush following load/elution)

PilotEvaluation

ProcessDesign

GapAnalysis


RunsEng

Runs

Development Runs & Scale-Up (cont.)• Optimize operations (cont.)

Confirm/adjust UF operating parameters– TMP vs. flux measurements at constant cross-flow– Minimize involvement of permeate PCV in TMP control

TMP vs. Permeate Flux

0

20

40

60

80

100120

140

160

180

200

220

2.0 4.0 6.0 8.0 10.0 12.0 14.0 16.0 18.0 20.0 22.0TMP (psi)

Per

mea

te F

lux

(LM

H)

0.0

10.0

20.0

30.0

40.0

50.0

60.0

70.0

80.0

90.0

100.0

Per

mea

te P

CV (%

open

)20kL scale TMP-fluxPermeate PCV

252 LPM

262 LPM

PilotEvaluation

ProcessDesign

GapAnalysis


RunsEng

Runs

Development Runs & Scale-Up (cont.)• Identify logic parameters that control operations

– Recognize and resolve code issues (transfers, load cell vs. flow meter)– Determine Delta V operating parameters (pump output %, PCV settings)

• Characterize process– Compare IP and release testing results to acceptance criteria– Perform AKTA “use” test with production buffers/supplies – Sample map for peak profiling and UF characterization

PilotEvaluation

ProcessDesign

GapAnalysis


RunsEng

Runs

Elution Overlays

Collection Volume (CV)

UV

Abs

. (O

D)

2.6cm Cell Sim2.0m DR3 2.0m ENG 4

End of collection

Elution Process Parameters

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0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 2.4 2.6 2.8 3.0 3.2 3.4Column Volumes (CV)

Flow

(L/m

in),

Col

umn

Inle

t (ps

i)

0.0

2.0

4.0

6.0

8.0

10.0

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20.0

UV

Abs

. OD

Pressure Control Valve

Outlet Flow (L/min)

Column Inlet Pressure

Outlet UV Meter 2

Development Runs & Scale-Up (cont.)• Process Modifications

Respond to unexpected process events– Disruption of elution operation at increased bed height – Placed in hold due to large ∆P across column

Elution Process Parameters

0

10

20

30

40

50

60

70

80

90

100

0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 2.4 2.6 2.8 3.0 3.2 3.4Column Volumes (CV)

Flow

(L/m

in),

Col

umn

Inle

t (ps

i)

0.0

2.0

4.0

6.0

8.0

10.0

12.0

14.0

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18.0

20.0

UV

Abs

. (O

D)

Pressure Control Valve

Outlet Flow (L/min)

Column Inlet Pressure

UV Abs. (OD)

Column Inlet Pressure (blue) 59psi

Column Inlet Pressure (blue) 37psi

PilotEvaluation

ProcessDesign

GapAnalysis


RunsEng

Runs

Issue Solution

Development Runs & Scale-Up (cont.)

• ConsiderationsDefine process as early as possibleDesign process for variety of fermentation outcomesConservative estimate of buffer usage (skid prime, line hold-up, ILD)Scheduled adequate time for wet-testing, documentation and validationDelta V controls studio on-line to troubleshoot code real-time

PilotEvaluation

ProcessDesign

GapAnalysis


RunsEng

Runs

Engineering Runs

• Aims and ObjectivesOperations compliant with cGMPManufacture comparable drug substanceAssess deviationsConfirm robust and reproducible process

Tighten ControlEnable alarm conditionsFine tune parameters to ensure full automation

PilotEvaluation

ProcessDesign

GapAnalysis


RunsEng

Runs

Engineering Runs (cont.)

• Demonstrate Reproducibility

PilotEvaluation

ProcessDesign

GapAnalysis


RunsEng

Runs

Eng Runs Elution Overlays

0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 2.4 2.6 2.8 3.0 3.2 3.4 3.6 3.8 4.0 4.2Column Volumes (CV)

UV

Abs

. (O

D)

Consistency & Commercialization

• Aims and ObjectivesDemonstrate lot to lot consistencyMeet Critical Process Parameter (CPP) criteriaMeet in-process and DS release specificationsAssess deviations

PilotEvaluation

ProcessDesign

GapAnalysis


RunsEng

Runs

Consistency & Commercialization (cont.)

• Deviation AssessmentComplex due to automated facilityRequires knowledge of in code, P&ID’s, operationsCase in point (UF over-concentration)– Goal: concentration target– Verify: calculation in phase logic – Phase end-point: totalized permeate volume removed– Assumption: target accounts for skid hold-up volume– Problem: membrane equilibration did not incorporate retentate

return line (field and P&ID verified)– Result: amount of over-concentration equals void volume– Solution: Include factor in BR to reduce permeate volume

target in logic

PilotEvaluation

ProcessDesign

GapAnalysis


RunsEng

Runs

Conclusions

• Rapid facility start-up and validation of multiple products– Start of Engineering Design: October 2000– Broke ground: September 2001– Pilot Run: March 2004– GMP Production: August 2004– First Approval to Manufacture: September 2005

• Success Required:– Careful attention to facility design– Pilot and scale-up models– Clear and decisive communication– Commitment from multi-disciplinary groups

• Important to have full customer interaction and support

Acknowledgements

• Jim Davies• Bruce McCafferty• Martin Smith• Ken Green• Carol Knevelman

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