1. A Tech Transfer Case Study From a CDMO
Sigma S. Mostafa
Director, Process Development KBI Biopharma

2. Overview Introduction Tech Transfer in a CDMO Case Study CDMO KBI
Unique constrains
KBI’s work paradigm
Case Study

3. What is a CDMO? 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
Confidential

4. Durham Site

5. KBI Locations KBI Durham Analytical and Formulation Dev.
Cell Line Development
Cell Culture cGMP Manufacturing
Microbial cGMP Manufacturing
KBI Boulder
Microbial Process Development
Microbial cGMP Manufacturing
Analytical Development
KBI RTP
Cell Culture Process Development
Downstream Process Development
Microbial Process Development
Analytical Development
Confidential

6. About KBI – Our Services
Cell line Development
Analytical Method Development
Preformulation and Formulation Development
Process Development
API Manufacturing
-Microbial
-Mammalian
Release testing and stability studies
Confidential

7. Types of Projects in Process Development
Process Transfer
Full Process Development
Material Supply
Process Characterization
Process Optimization

8. 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

9. Process Development Work Flow
- 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

10. Tools of Tech Transfer
Development Report, Demo Report, Process Description
Raw Data
Process Control Trends
Process Flow Diagrams
Bill of Material
Risk Assessment

11. Technology Transfer – Key Contributors
30+ with advanced scientific and technical degrees in Proc Dev
30+ manufacturing staff with significant operations experience with small and large biotech/biopharma
People:
experienced &
dedicated
Process: well characterized for scale up & Mfg
Facility
cGMP compliant
Facility fit
Program management: System for information
transfer
Confidential

12. KBI’s Business Process for Tech TransferPD
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
Method Development
Method Qualifications
Formulation Development
Sampling and Testing plans,
Specifications
IP and release
Stability
Facility Fit
Draft 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
Confidential

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

14. KBI’s Cell Culture Platform Process
Shake Flasks
ambrTM Bioreactors
3L Bioreactors
Medium and Supplement Screening
Process Parameter Screening
200L Bioreactor
3L Bioreactors
15L Bioreactors
Process Optimization and Robustness
Demonstration Run
Confidential

15. Mixing Characteristics in Bioreactors
Data from mixing studies used to set agitation rate,
aeration strategy, process control strategy

16. Comparability Across Scale – 3L, 15L, 200L, and 2000L
Viable Cell Density
VCD data matches across scale
 Gottschalk, U., Shukla, A. Single-use disposable technologies for biopharmaceutical manufacturing, Trends in Biotechnology, 31(3), , 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.

17. Comparability Across Scale – 3L, 15L, 200L, and 2000L
Titer
Titer data matches across scale

18. Scale-up studies
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
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), , 2014.

19. 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

20. 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.
Sep 2013
Tech Transfer to KBI (Shake flask and 3L bioreactor scale)
Material Supply Run
(200 L Scale)
Oct 2013
Process Optimization
(Ambr Study)
Nov 2013
Material Supply Run
(15 L Scale)
Process Optimization (3 L bioreactor-scale)
Dec 2013
Process Confirmation (15 L bioreactor-scale)
Jan 2014
Tech Transfer
Demo Run
(200 L-scale)
Feb 2014
cGMP Run
(2000 L-scale)
Mar 2014

21. 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

22. Case Study – Tech Transfer into KBI
At 3L bioreactor scale productivity with the
KBI fed-batch process was 2.2x higher than the
original batch process

23. 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 of
Process development
Initial cell growth and peak cell density in the
200L run was slightly higher than the 3L scale
Viability drop was faster in the 200L scale
Compared to the 3L fed-batch culture

24. Case Study – 200L Material Supply Run
Titer in the 200L scale did not match the
3L fed-batch data
A significant amount of base addition
occurred in the run
Maintaining pH within deadband (0.05 pH)
Was difficult

25. Case Study – 200L Material Supply Run
Glucose uptake was somewhat higher in the 200L
scale 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)

26. Case Study – Shake Flask & ambr Studies
Titer
Lactate Comparison
Shake flasks
ambr
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

27. 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

28. cGMP Manufacturing (2000 L-scale) Comparison w/ Small-scale
Cell growth and viability compared well among 3L, 200L, and 2000L scales
Vesper - Confidential

29. cGMP Manufacturing (2000 L-scale) Comparison w/ Small-scale
Lactate profile was much improved compared
to the 200L material supply run
Maximum lactate level in the 2000L was
4 g/L, less than half of the level observed in the
material supply run
Vesper - Confidential

30. cGMP Manufacturing (2000 L-scale) Comparison w/ Small-scale
Titer was comparable across 3L,
200L, and 2000L scales
4.5X increase in titer compared to
the 200L material supply run.
Vesper - Confidential

31. 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.

32. Acknowledgements Analytical Executive Management Tech Transfer
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