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Process Design for an All Single-Use Manufacturing Facility:Scaling Low to High Titer Processes to Fit Standard mAbEquipment

BioProcess International WestMarch 2, 2017

Kelly ThomAssociate Principal ScientistFujifilm Diosynth Biotechnologies

One Global Company

3SITES

Billingham, UKCollege Station, TXRTP, North Carolina

1,100EMPLOYEES

World Wide6LICENSES

For commercial manufacturing.

20+YEARS

Of Biologics CDMO experience.

280+MOLECULES

In process development and/or manufacturing.

With you all along the road To clinical success

Regulatory

ApprovalLaunchPhase IIIPhase I Phase IIPreclinical

Gene Expression &Strain / Cell Line

Development

Process Invention

Pre-clinicalManufacture

Process Development& Optimization

Analytical & Stability

cGMP ManufactureFill/Finish

ProcessCharacterization

Process Validation

cGMP Manufacture

Stability

CommercialProduction

Post-approvalActivities

with

55CHO Programs

Our Cell Culture Experience

65+Cell Culture Programs, including CHO

and Baculovirus

10Baculovirus Programs

1Commercially Approved

Cell Culture Product Manufactured at FDB

FDB Single-Use Bioprocessing Journey

• Learning from Early Adoption of Single-Use Technologies

• Process Design Goals for New Facility

• Facility Layout, Room Classifications, and Closure Strategy

• Upstream and Downstream Single-Use Processes and Equipment

• Modeling to Design Small (2 g/L), Medium (5 g/L), and Large (8 g/L) Downstream Process

Creating biomanufacturing capacityA global partnership

1000 L project

1000 L project

2000 L project

• Single-use WAVE Bioreactor™ systems

• Xcellerex™ XDR cell culture suite

• ÄKTA™ready and ReadyToProcess™

prepacked columns

2013

2012

2014

2015

Billingham, UK

Driven by the industry need for

cGMP mammalian cell culture

biomanufacturing capacity

Four successful capacity expansion

projects on two continents

Creating the UK’s first fully single-use

biomanufacturing facility

2017 20182019

• Xcellerex single-use mixers

• Xcellerex XDR cell culture suite

• ÄKTA™ready purification system

• Xcellerex XDR 2000L

bioreactor added

Research Triangle Park, US

Research Triangle Park, US

2000 L project

• Xcellerex XDR 2000L

bioreactor added

Billingham, UK

Ongoing partnership

Development of FDB Single Use Bioreactor Platform

• Why Single Use?– Capacity can be increased quickly– Capital investment and payback time are less– Footprints are reduced– Less utilities are required (CIP/SIP)– No cleaning verification/validation– Less cost for room classifications, EM monitoring, personnel gowning/flow– Multi-product manufacture (“ballroom”) suites– Multiple products in multiple suites (reduced changeover times)

• Production Bioreactor Selection– Process performance (mixing, mass transfer, sparger flexibility)– Engineering design (hardware and bag design)– Control system capability and automation tie-in– Equipment cost and product support– Speed and ease of implementation and qualification– Track record

200L SUB

1000L SUB

110L SS

0

50

100

150

200

250

300

0 2 4 6 8 10 12 14 16 18 20

VC

D (

10

5c

ell

s/m

L)

Time (days)

110LSS Avg (n=3) 200LSUB Avg (n=3) 1000L SUB Avg (n=2)

Stainless Steel vs. Single Use Bioreactor

0

200

400

600

800

1000

1200

0 2 4 6 8 10 12 14 16 18 20P

rod

uc

t (g

/L)

Time (days)

110LSS Avg (n=3) 200L SUB Avg (n=3) 1000LSUB Avg (n=2)

Product Quality: Glycosylation Patterns

SUB Scale Up by Mass Transfer

• O2 kLa characterization performed for all SUBs– 10, 50, 200, 500, 1000 and 2000 L

• kLa models are being developed in MODDE

• CO2 stripping characterization is on the horizon

± 20%

0 – 50%

Dis

so

lve

d O

xyg

en

(%

) O2

Sp

arg

e(s

Lp

m)

Culture Duration (14 days)

1000 L CHO Process (~30 E6 cells/mL)

To achieve peak kLa = 7 hr-1

100 rpm = 50% sparge120 rpm = 38% sparge

SUB Scale Up to 1000 L GMP

Upstream Learning from Early Adoption

• Hardware Design– MFC sizing

– Automated exhaust filter strategy

• SUB Bag Design– Enough addition lines with correct

diameters

– Tubing is long enough for all connections

– Multiple exhaust filters

– Evaluate the bag film

• Connectivity– Minimize the number of sterile

connections

– Welding vs. sterile connectors

– Solution bags with on-board filters and tubing to match the SUB

• Raw Materials– Liquid media

– Pre-made solutions (nutrient feeds if stability allows, glucose, glutamine, antifoam)

– Have capability to formulate if needed

Development of FDB Single Use Downstream Platform

• Manufacturing experience with multiple single-use vendors and equipment

Chrom Skids Pre-packed Columns Single Use TFF Skids Single Use Mixers

GE AKTAReady GE ReadytoProcess Pall SU TFF GE XDM / XDUO

Repligen Opus Sartorius FlexAct Sartorius Palletank

Millipore Mobius

Downstream Learning from Early Adoption

• Design Limitations– Skids: flow rates, mixing, temperature control, hydroxide exposure,

flow kit max usage time (limited tubing lifetime in peristaltic pump)

– mAb throughputs: low chrom skid flow rates, low TFF membrane throughput

• Instrumentation Limitations– Sensor (P, T, UV, flow, conductivity) issues

– Use of traditional pH probe instead of inline pH probe

• Installation Limitations– Operators must standardize sensors as part of self check

– Manifold installation is cumbersome

– Lack of manifold labeling can result in cross connectivity issues

– Standard manifold tubing sizes may require different connectors (connecting 1” to ½” tubing)

• Tubing management and consumable design were key early learnings

• Need to manage customization against cost

Process Design Goals for HT, mAb Facility

• Standardized offering • All single-use equipment • Medium/high density fed batch CHO (10 – 40 E6 cells/mL)• Downstream to process a wide range of upstream titers

– Small (2 g/L), Medium (5 g/L) and Large (8 g/L) Downstream Scenarios

• One set of standard equipment with scaling flexibility• Define closed processing needs and connectivity strategy• Placement of unit operations for optimal suite scheduling and process flow• Allowable process duration for each unit operation• Off-the-shelf products wherever possible • BOMs for small, medium, and large downstream• Raw material and consumable costs for a batch

Design Approaches

• GE Flex Factory equipment and automation (USP, partial DSP)

• Super Pro Modeling: Created a simulation to scale the FDB platform process over a range of 2 – 8 g/L

Model Inputs Model Outputs

2000 L Harvest Titer Unit Op and Total Batch Duration

Filter Fluxes Column Sizes

Resin Capacities Number of Column Cycles

Step Yields Filter/Membrane Areas

Buffer and Process Volumes

SUM Sizes

Raw Material Cost per Gram

Unit Operations by Suite

• Harvest one batch per week• At steady state, up to nine products in flight

Single-Use Upstream PFD

Single-Use Downstream PFD

Super Pro Model

Protein A ViralInactivation

CEX

AEX Nanofiltration TFF Bulk Fill

Chromatography Estimates and SUM Sizing

Protein A CEX

Protein Titer Column Size CyclesProcess

DurationColumn Size Cycles

Process Duration

(g/L) (L) (#) (hr) (L) (#) (hr)Small 2 - 3 10 8 - 12 48 - 62 10 6 - 9 20 - 30Medium 3 - 5 20 6 - 10 31 - 45 20 4 - 7 14 - 24Large 5 - 8 32 7 -10 31 - 41 32 5 - 7 17 - 24

Clarified Harvest

VI SUM 1 VI SUM 2 CEX Eluate AEX Eluate Nanofiltrate

Protein Titer SUM A SUM B SUM C SUM D SUM E SUM F

(g/L) (L) (L) (L) (L) (L) (L)Small 2 - 3 2500 200 200 1000 1000 1000Medium 3 - 5 2500 500 500 1000 1000 1000Large 5 - 8 2500 1000 1000 2500 2500 1000

NOTE: SUM sizes were determined from process volumes, which are not shown.

Raw Materials Cost per Gram

TFF Design

Protein Titer (g/L)

DS Conc. 2 3 5 8

(mg/mL) TFF Retentate / Final DS Volume (L)

1

10 236 354 590 944

50 47 71 118 189

100 24 35 59 94

180 20 33 52

• Assumption that DF will occur at 10 – 50 mg/mL retentate concentration• 50 L and 200 L retentate tanks will be used• 1000 L SUM used for retentate volume > 200 L

Summary

• FDB leveraged learnings from early adoption of single-use technologies to guide process/facility design

• The new facility utilizes the multi-product ballroom approach– Harvest of one batch per week

– Up to nine products in flight at steady state

• Super Pro modeling was used to define small, medium and large downstream processes– Equipment

– Processes

– Durations

– Cost

Acknowledgements

Upstream Development

• Sharyn Farnsworth

• Simon Uphill

Downstream Development

• Patrick Daley

• Mark Chavez

• Jonathan Haigh

• Michael Murray

• Phil Ropp

• Kevin Short

• Matt Teten

Process/Facility Design

• Mike Jones

• Peter Large

• Stewart McNaull

• Thomas Page

• Mary Vo-Harris

• GE Flex Factory Design Team

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