Next Generation Processes: What Model Works Best to Manufacture Recombinant Proteins in Asia? BioPharma Asia 2017 Suntec Convention Center. Singapore, March 22, 2017 Thomas Jung, M.S. Vice President, Business Development KBI Biopharma Inc.

1. Next Generation Processes: What Model
Works Best to Manufacture Recombinant
Proteins in Asia?
Thomas Jung, M.S.
Vice President, Business Development
KBI Biopharma Inc.
BioPharma Asia 2017
Suntec Convention
Center
Singapore
March 22, 2017

2. Internal Manufacturing vs CMO

3. Internal vs CMO Considerations
Molecule
Indication
Target Market(s)
Manufacturing Yield
Facilities Cost
Access to Expertise
Location of CMO

4. Type of Molecule
• Innovator vs biosimilar
• Therapeutic antibody, recombinant protein,
antigen, peptide
• Stage of development: preclinical, early
clinical, late stage, commercial
• Indication
• Route of administration
• Dose: known or estimated
• Dosage will influence manufacturing
demand.

5. Target Market(s)
• Selected country or countries, region, or
global?
• Regulatory requirements may influence
manufacturing strategy.
• Clinical / commercial drug substance / drug
product requirements.
• Market scope will influence DS / DP
requirements and manufacturing demand.

6. Manufacturing Yield
• Production clone developed?
• RCB Available?
• Yield of current cell line adequate?
• Consider development of higher yielding cell
line.
• Higher yielding cell line reduces
manufacturing scale.
• Higher productivity cell lines / more costly
licensing terms.

7. Internal Manufacturing
Considerations
• Costs
• Facility cost
• Equipment cost
• Development and manufacturing personnel cost
• Quality systems cost
• Development Strategy
• Stage of development
• Exit strategy: Sell asset? At what phase?
• Partner with large pharma, biotech?
• Risk
• Postpone internal manufacturing costs?
• Build internal manufacturing later, if needed?

8. CMO Option
• Advantages of Using CMO
• Contract vs build (rent vs buy)
• Access to existing facilities
• Access to CMO expertise
• Selection Criteria
• Strong and relevant experience
• Scientific expertise
• Implementation of latest innovations
• Quality systems / regulatory history
• Program management
• Geographic location of CMO
• Close proximity not essential
• Good communications are critical

9. Single Use Bioreactor vs
Stainless Steel

10. SUB vs SS Considerations
Scale of Manufacturing
Quality
Flexibility
SUB vs SS Comparison

11. Scale of Manufacturing
*Yields are based on upstream expression levels.
with downstream recoveries assumed to be 70%.

12. SUB Quality
Single Use Bioreactors:
• No CIP/SIP reduces changeover
time.
• Significantly decreases possibility of
product carryover or cross-
contamination from one campaign
to the next.
• Increased facility throughput.
• Reduced timelines for sequential
batch manufacturing.
• Flexible manufacturing scale
matches process scale to DS
requirements, reducing materials
costs.

13. SUB Flexibility
• Flexible manufacturing scale of GE xcellerex SUB with
a 5:1 turndown ratio – 2000L SUB working volume
scalable from 400L to 2000L.
• SUB maximum working volume of 2000L could be
limitation.
• Multiple DS batches at 2000L may satisfy DS
requirements – e.g. 2 X 2000L and possible pooling
for purification as single batch.
• SUB shows good comparability to SS such that
transfer to larger scale SS bioreactor can be made for
late stage / commercial, if needed.

14. SUB vs Stainless Steel
Single Use Facility Relative to Stainless Steel
Water Usage 87% reduction
Cleaning Chemicals Usage 95% reduction
Energy (Electricity) Demand 30% reduction
Facility Footprint 38% less
Steelwork 62% less
Headcount 21% lower
Plastic Waste 880 kg increase
CO2 Emissions 26% reduction
Comparison based on 3 X 2000L MAb Production Scale*
* The Environmental Impact of Disposable Technologies
By Andrew Sinclair, Lindsay Leveen,Miriam Monge,Janice Lim,Stacey Cox

15. Accelerated Manufacturing
Timelines

16. Time is of the Essence
Bring innovative therapies to patients sooner.
Innovative biologic drugs improve patient survival,
quality of care, and quality of life.
Intense competition for share of biosimilar market
requires early approval and entry into market to be
successful.
High cost of drug development requires shortening time
to return on investment for venture capitalists and
investors.
Need to exploit any opportunities to shorten clinical trial
/ regulatory approval process without sacrificing quality.

17. Opportunities to Accelerate
Cell Line Development
Cell Culture Process Development
Purification Process Development
Analytical Method Development
Drug Product Formulation Development
Tox Manufacturing

18. Cell Line Development
Technologies
Vector
Construction
Transfection &
Selection
(Amplification)
Pool
Enrichment
Clone Isolation
& Screening
Stability
Assessment
Strong vector w/
enhance element
Transient
Transfection
Gene
Stable
Pool
FACS
High throughput
cloning
(FACS or
ClonePix)
Gene codon
optimization Shaker
24-wp
clone
screening
CSI
Process
Development
Process
Development
Cell
Bank
High Throughput clone selection (Clonepix, FACS) screens larger
number of clones and selects clones based on productivity
compared with traditional limited dilution cloning.

19. Shaker 24-Well Plate Clone
Screening
10 days
14‐21 days
10 days
Static culture
Shaker culture
(deep 24‐wp)
10 days
14‐21 days
0
5
10
15
20
25
30
35
40
45
0 5 10 15 20 25 30 35 40CloneRankingbyd24wp
Clone Ranking in SF 7 day batch culture
New cloning screening process reduces timeline by 10 days.
Shaker culture is introduced as early as possible, so that the
clones screened out fit into the downstream scale up model.
Good correlation between two steps

20. High Throughput Cell Culture
Process Development
Basal medium, feed medium, and process parameter studies
can be performed at 9-16 ml scale rather than 2L or 10L
working volume. Conventional process development timelines
of 3-4 months can be reduced to 1-1.5 months using ambr.
1mL tips
1mL
or
4mL tips
Two culture stations; each holding 12 bioreactors.
All 24 bioreactors independently controlled for pH and DO.
Used Tips
Discard
Liquid
Handler
ambrTM Technology
Microbioreactor
(9-16mL working volume)

21. Scale Up Studies
Cell Growth Titers Product Quality Attributes
• 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.

22. Platform mAb Purification Process
Bulk Fill
UF/DF: formulation buffer exchange, concentration adjustment
Virus filtration
Chromatography (can be flow through)
Purification IEX chromatography
Low pH viral inactivation
Protein A capture chromatography
Cell separation by depth filtration

23. High Throughput Analytics
LabChip GXII has
capability to run specific
assays for Protein Glycan,
Protein Charge Variant
(CZE), and Protein
Molecular Weight
Octet instrument is used
for quick turnaround ProA-
based titer analysis of
diluted, high concentration,
in process cell culture
samples.
High Throughput Analytics allow for rapid titer
determinations and assessment of critical quality attributes,
leading to faster decisions and reduced timelines in cell line
development and process development.

24. Analytical – Utilize Platform
Methods as Appropriate
• Protein Primary Structure
 Peptide Sequencing via LC/MS/MS
 Amino Acid Analysis
 Peptide Mapping
• Biophysical Characterization
 CD, FTIR, DSC, DLS, fluorescence
spectroscopy
• Capillary and Slab Gel Electrophoresis
 CZE
 SDS-CGE
 cIEF and icIEF
 SDS-PAGE and IEF
 Western blot
 Microchip electrophoresis
 2D gels and blots
• Glycan Analysis
 Oligosaccharide mapping
 Monosaccharide composition
 Sialic Acid Quantitation
• Process Residuals
• ELISA (HCP, protein A etc.)
• HPLC (antibiotics, IPTG, detergents, etc)
• qPCR (DNA)
• HPLC
• Size Exclusion (with MALLS)
• Ion Exchange
• Reverse Phase
• Hydrophobic Interaction
• Affinity
• Potency Assays
• Binding Assays via ELISA, Biacore and
ForteBio
• Cell Based Assays (e.g., proliferation,
cytokine release, etc.)
• Mass Spectrometry
• Intact mass
• Peptide mapping with LC/MS or
LC/MS/MS
• Disulfide Mapping
• Post translational modifications (e.g.,
oxidation, deamidation)
• PEGylation site identification
• Glycan Identification & site identification

25. Preformulation: Design of Experiments (DOE)
30-40 Candidate
Formulations:
Various buffer, pH,
ionic strength, and
excipient conditions
Biophysical Screening: DSC,
DLS, CD, FTIR, Fluorescence
Select Candidate
formulation(s)
with appropriate
thermal and
structural stability
High Throughput Preformulation screening approach selects optimal
formulations to carry into forced degradation and final dosage development
based on biophysical characteristics and guided by statistical analysis.

26. Platform Expansion Process
Wave 20/50
Vial Thaw
Xcellerex 2000L
Xcellerex 200L
Shake Flask
Expansion
Seed or Production Bioreactor
Production Bioreactor
Each passage 2 – 4 days
Production 2 – 3 weeks

27. Production Options for Preclinical
Tox Drug Substance Production
Conservative Approach
• Engineering run performed in
cGMP suite
• Same single use bioreactor
platform as PD
• Scale up to 2000L
• Source of reference material
• Viral clearance sampling
• More control
• Less risk
• Traditional timeline
• Slot reservation considerations
Accelerated Approach
• Demonstration run performed
in process development suite
• Same single use bioreactor as
cGMP
• Scale limited 200L
• Source of reference material
• Viral clearance sampling
• Less control
• More risk
• Shorter timeline
• More flexibility in scheduling

28. Conclusions
• Using a CMO for production of early clinical
drug substance offers a compelling value
proposition.
• Employing Single Use Bioreactors provides
a strong quality advantage and maintains
flexibility in terms of clinical and commercial
manufacturing.
• Reducing timelines through effective use of
technology innovations and manufacturing
strategies may shorten time to approval and
increase return on investment.

29. Thank you!