A Tech Transfer Case Study From a CDMOSigma S. MostafaDirector, Process DevelopmentKBI Biopharma10-03-2015

Overview• Introduction

– CDMO– KBI

• Tech Transfer in a CDMO– Unique constrains– KBI’s work paradigm

• Case Study

What is a CDMO?

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• Contract Development & Manufacturing Organization

• Responsible for clinical trial material (CTM) development and manufacturing– Process development & manufacturing of Bulk Drug

Substance (BDS)– Formulation and manufacturing of dosage forms, i.e.

Drug Product (DP)– Analytical support for BDS and DP

• A large portion of CTM development & manufacturing expenditure is outsourced to CDMOs

a customer and science-focused contract development & manufacturing organization

DURHAM SITE

KBI Locations

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KBI RTP• Cell Culture Process Development• Downstream Process Development• Microbial Process Development• Analytical Development

KBI Boulder• Microbial Process Development• Microbial cGMP Manufacturing• Analytical Development

KBI Durham• Analytical and Formulation Dev.• Cell Line Development• Cell Culture cGMP Manufacturing• Microbial cGMP Manufacturing

About KBI – Our Services

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Cell line Development

Analytical Method

Development

Preformulation and

Formulation Development

Process Development

Release testing and

stability studies

Types of Projects in Process Development

Process Transfer

Full Process Development

Material Supply

Process Characterization

Process Optimization

Tech Transfer Aspects in a CDMO• Large number of tech transfers per year

– KBI has >10 tech transfers per year from PD to manufacturing

• Each cell type, molecule, and process are different; opportunities to leverage platform is limited

• Timeline for tech transfer is short and overlaps with development work– PD scale-up run and first manufacturing run are 1 -2

months apart– Short timeframe necessitates staggered tech transfer

approach

Process Development Work Flow

Process Development

- Kick-off meeting- Shake flask study- ambr, 3L, & 15L bioreactor study- Harvest study

Tech Transfer to Manufacturing

- Facility Fit- Risk Assessment- BOM- PFD- Process Description

Scale-up Run

- 200L Disposable Reactor- MCB vial- Representative seed train- Final Process

Tools of Tech Transfer

Development Report, Demo Report, Process Description

Process Flow Diagrams Bill of Material Risk Assessment

Raw Data Process Control Trends

Technology Transfer – Key Contributors

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Process: well characterized for scale up &

Mfg

FacilitycGMP compliant

Facility fit

Program management: System

for information transfer

People: experienced &

dedicated

• 30+ with advanced scientific and technical degrees in Proc Dev

• 30+ manufacturing staff with significant operations experience with small and large biotech/biopharma

KBI’s Business Process for Tech Transfer

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PD

AFS

MFG

QA

Client

Develop upstream and downstream

Perform Confirmation and Demo runs

Prepare process overview, facility fit, process description, solution and

buffer recipes, sampling plan w/AD

Review batch records, person in plant for key steps

for 1st run

Facility FitDraft batch records

and solution records

Finalize BOM, order raw materials, ensure solutions and buffers specifications

Execute Eng and GMP runs, close

deviations

Campaign Summary

Review and approve MBRs, solutions

records, sampling plans

Deviation closure and Batch release

Review process overview, facility fit and process

descriptions

Approve process descriptions, BRs, BOM, solutions records,

person in plant as decided

Method Development

Method Qualifications

Formulation DevelopmentSampling and Testing plans,Specifications

IP and releaseStability

Single use disposable technologies

Media and Feed preparation utilizing disposable mixing, filtration and storage systems

Disposable shake flasks or disposable spinner flasks

MCB or WCB vial

Disposable expansion reactor

Disposable seed bioreactor

Disposable production bioreactor

Disposable fluid path centrifuge

Disposable depth filtration system

0,2 µm filter

Hold vessels (Bags)

Hold vessel (bag)

Disposable fluid path purification system

Disposable mixing tank

0,2 µm filter

Retentate

Permeate

PD

Disposable fluid path purification system

Disposable mixing tank

0,2 µm filter

BPC

Virus filter

BPC

0,2 µm filter

BPCBPC

Sterile bulk fill and sampling bags

Buffer preparation utilizing disposable mixing, filtration and storage systems

0,2 µm filter

Disposable fluid path UF/DF system

Aseptic connection

Hold vessel (bag)

Hold vessel (bag)

Hold vessel (bag)

Hold vessel (bag)

Hold vessel (bag)

  Shukla, A., Mostafa, S., Wilson, M., Lange, D. Vertical Integration of Disposables in Biopharmaceutical Drug Substance Manufacturing, Bioprocess International, 10(6), 34-47, 2012.

KBI’s Cell Culture Platform Process

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Medium and Supplement Screening

ambrTM Bioreactors 3L Bioreactors

Shake Flasks

Process Parameter Screening

Process Optimization and Robustness

3L Bioreactors 15L Bioreactors

Demonstration Run

200L Bioreactor

Data from mixing studies used to set agitation rate, aeration strategy, process control strategy

Mixing Characteristics in Bioreactors

Viable Cell Density

• VCD data matches across scale

Comparability Across Scale – 3L, 15L, 200L, and 2000L

 Gottschalk, U., Shukla, A. Single-use disposable technologies for biopharmaceutical manufacturing, Trends in Biotechnology, 31(3), 147-154, 2013. Shukla, A., Mostafa, S., Wilson, M., Lange, D. Vertical Integration of Disposables in Biopharmaceutical Drug Substance Manufacturing, Bioprocess International, 10(6), 34-47, 2012.

• Titer data matches across scale

Titer

Comparability Across Scale – 3L, 15L, 200L, and 2000L

• Comparison across scales for the production of a recombinant glycoprotein in a recombinant CHO cell line

– The process decisions and results from ambrTM were reproducible to other scales

Scale-up studies

Cell Growth Titers Product Quality Attributes

Rameez, S., Mostafa, S., Miller, C., Shukla, A. High-throughput miniaturized bioreactors for cell culture process development – reproducibility, scalability and control, Biotechnology Progress, 30(3), 718-727, 2014.

Case Study – Project Start-up

• Expedited Process Development for a novel mAb

• Data from client on shake flask batch study• CHO DG44 cell line from client (prepared by a

third party)• Client expressed need for 200L scale material

delivery before process development started

Case Study – Project Scope of Work• Contract Signed in early September• Face to Face Kick-off meeting in mid September• Tech Transfer into KBI – A shake flask study and a 3

x 3L bioreactor study completed by mid October• A 200L Disposable Bioreactor Supply Run

completed by mid October • Studies at ambr, 3L and 15L bioreactor scaled

conducted November through January• PD Demonstration Run at 200L Disposable

Bioreactor done in Feb• Vial thaw for cGMP run in KBI manufacturing

facility in March• Start of project to manufacturing vial thaw in 7

months for a full development mAb project.

Tech Transfer to KBI (Shake flask and 3L

bioreactor scale)

Process Optimization (Ambr Study)

Process Confirmation (15 L bioreactor-scale)

Process Optimization (3 L bioreactor-scale)

Demo Run (200 L-scale)

cGMP Run (2000 L-scale)

Material Supply Run(15 L Scale)

Material Supply Run(200 L Scale)

Sep 2013

Oct 2013

Nov 2013

Dec 2013

Jan 2014

Feb 2014

Mar 2014

Tech Transfer

Case Study – Tech Transfer into KBI

• Client transferred shake flask batch process• In Tech Transfer Run, client’s process was carried out in shake flask and 3L reactors• In parallel one feed was tested in preparation for 200L material supply run• Cell growth improved in the fed-batch culture

Case Study – Tech Transfer into KBI

• At 3L bioreactor scale productivity with theKBI fed-batch process was 2.2x higher than theoriginal batch process

Case Study – 200L Material Supply Run

• An initial 200L Material Supply Run was carried out following the 3L Tech Transfer Run• This 200L run was done prior to initiation ofProcess development• Initial cell growth and peak cell density in the200L run was slightly higher than the 3L scale• Viability drop was faster in the 200L scaleCompared to the 3L fed-batch culture

Case Study – 200L Material Supply Run

• Titer in the 200L scale did not match the3L fed-batch data

• A significant amount of base additionoccurred in the run

• Maintaining pH within deadband (0.05 pH)Was difficult

Case Study – 200L Material Supply Run

• Glucose uptake was somewhat higher in the 200Lscale compared to the 3L fed-batch culture

• Final lactate level was around 10 g/L in the 200L scale (compared to 7 g/L in 3L fed-batch reactor)

Case Study – Shake Flask & ambr Studies

Design-Expert® Software

Correlation: 0.331Color points by level ofA:Feed

Feed A+BFeed (A+B)Feed CCell Boost 6

Ha

rve

st

Tit

er

(Da

y 1

2 o

r D

ay

14

)

Feed A+B Feed (A+B) Feed C Cell Boost 6

0.2

0.29

0.38

0.47

0.56

• Project required expedited process development• Shake flask study focused on feed and supplement evaluation• ambr study focused on pH set-point and feed impact on lactate & titer

Tite

r

0.00

0.50

1.00

1.50

2.00

2.50

4 5 6 7 8 9 10 11 12 13 14

Lact

ate

(g/L

)

Time (Days)

CS1 - Control (pH 7.0)CS2 - Control (pH 7.0)Cell Boost 6 - pH 6.90Feed C - pH 6.90

Lactate Comparison ambrShake flasks

Case Study – 3L Bioreactor Study

• A fractional factorial DOE was carried out in the 3L scale• Multiple feeds, temperature scheme, and pH set point were tried• pH dead band was expanded• Feeds with lower lactate level and higher productivity were identified

Vesper - Confidential

cGMP Manufacturing (2000 L-scale) Comparison w/ Small-scale

• Cell growth and viability compared well among 3L, 200L, and 2000L scales

0

2

4

6

8

10

12

14

16

0 2 4 6 8 10 12 14 16

VCD

(1 x

10^

6 ce

lls/m

L)

Time (Days)

2000 L GMP Run #12000 L GMP Run #2200 L-scale PD Demo Run3 L-scale Final Process

0.0

20.0

40.0

60.0

80.0

100.0

120.0

0 2 4 6 8 10 12 14 16

Cell

Via

bilit

y (%

)Time (Days)

2000 L GMP Run #12000 L GMP Run #2200 L-scale PD Demo Run3 L-scale Final Process

Vesper - Confidential

cGMP Manufacturing (2000 L-scale) Comparison w/ Small-scale

• Lactate profile was much improved comparedto the 200L material supply run

• Maximum lactate level in the 2000L was4 g/L, less than half of the level observed in thematerial supply run

Vesper - Confidential

cGMP Manufacturing (2000 L-scale) Comparison w/ Small-scale

• Titer was comparable across 3L,200L, and 2000L scales

• 4.5X increase in titer compared tothe 200L material supply run.

Conclusions• Understanding of cell line characteristics and process parameter impact on

cell line is imperative for successful tech transfer• Use of high throughput systems such as ambr micro-bioreactors provide significant advantage during expedited process development• For an expedited manufacturing plan, a phased approach to tech transfer

is needed; identifying and ordering the long lead items and determining facility fit are often the most rate limiting activities

• Testing of scalability early in process development allows identification of cell line specific scalability challenges; therefore, using material supply runs as scale-up tests is advisable.

Acknowledgements

Process Development- Niket Bubna- Lynwel Cunanan- Brian Baker- Ronnie Nichols

Tech Transfer- Sam Pallerla

-

Manufacturing- Les Smith- Michael Huerta- Joaquin Lopez

Analytical

- Michael Pollock- James Smedley

Executive Management- Abhinav Shukla- Prathima Acharya- Joe McMahon