Abhinav A. Shukla, Ph.D. Senior Vice President, Development & Manufacturing KBI Biopharma, Durham NC & Chief Scientific Officer, JSR Life Sciences (Bioprocess) Biopharma India 2017, Sept 19-20th, Mumbai, India

1. Highly accelerated platforms
for mAb and next generation
mAb manufacturing
Abhinav A. Shukla, Ph.D.
Senior Vice President
Development & Manufacturing
KBI Biopharma, Durham NC
&
Chief Scientific Officer
JSR Life Sciences (Bioprocess)
Sunnyvale CA
Biopharma India 2017, Sept 19-20th, Mumbai, India

2. 2
What is KBI Biopharma
• Contract Development & Manufacturing Organization for biologics
• Based out of Durham, NC
• Started in 1997 as a technology company
• Initiated Analytical Services in 2004
• Initiated Process Development & Manufacturing in 2010
• Strong emphasis on Process Development expertise across all
modalities of biologics from entering the clinic to commercial launch
 12-13 INDs filed every year
 4 commercial process development programs
 2-3 standalone PC/PV programs for big pharma
 Microbial commercial launch (PPQ complete)
 Mammalian commercial launch (PPQ in 2018)

3. Confidential
3
Where We Are- Global Presence
Geneva
Leuven
Durham
RTP
Boulder
San Diego
The Woodlands
KBI Offices

4. 4
What is JSR Life Sciences
• Parent company of KBI Biopharma since 2015
• Business unit of JSR Corporation (Tokyo, Japan)
• > 50 year tradition in polymer innovation & manufacturing
• Life Sciences division is based out of Sunnyvale, CA
• Sites in Tokyo, Tsukuba (Japan), Leuven (Belgium) and
Beijing (China)
• Key emphasis on:
• In vitro diagnostics (MBL Laboratories)
• Contract Development & Manufacturing (KBI Biopharma)
• Bioprocess products for the Life Sciences industry e.g. Amsphere
A3 Protein A chromatography resin

5. Selexis – latest part of the KBI/JSR family
• Leading independent cell line in biotech industry
• Based in Geneva, Switzerland
• > 5 g/L titers for mAbs
• Significant track record with hard to express proteins
• Selexis Genetic Elements enable enhanced transcription via high
expression vectors

6. • mAb development from gene to GMP in ~ 9 months
• Single cycle of development
6
Pre-Clinical Phase I Phase II Phase III
Process Development
Process
Characterization
Process
Validation
Process Monitoring &
Improvement
FIH Process
• Deliver clinical process
quickly
• Platform process
• Clinical Supply
Submission &
Approval
Lifecycle
management
BLA Prep &
PAI
Commercial Process
• Deliver manufacturing process for
registrational trials and market
• Design keeping large-scale manufacturing
in mind
• Improve productivity, efficiency, robustness,
manufacturability, COGs
• Analytical characterization and method
development
Process Characterization and Validation
• Develop IPC strategy through understanding of process inputs and
outputs (design space)
• Scale-down characterization and validation studies
• Large-scale process validation to demonstrate process consistency
• BLA preparation
• Supporting documents for licensure inspections
• Post-commercial process improvements (CI)
• Post-commercial process monitoring
FIH process Commercial process

7. 7
Improvements in platform technology can enable
one process development cycle
• Essential for biosimilars or highly accelerated
programs
• Streamlined process
characterization/validation effort
Process
Characterization
Lifecycle
Management
Platform Application
IND BLA Commercial
Platform Technology Development

8. 8
Outline
• Where are mAb platforms today?
• Is there a universal downstream platform?
• Can platform approaches be extended for next generation mAbs?
• Can a platform approach be taken for commercialization?
• Process characterization studies leading to definition of an in-
process control strategy (IPC) – how fast can these be
completed?
• How innovation makes a difference in COGS

9. 9
Fundamental understanding of
bioprocesses is key
• Robust platforms can only be developed if there is a
strong understanding of the science of developing
bioprocesses
• Advanced platforms that are universal
• Multimodal chromatography
• Improved Protein A resins
• High Throughput Process Development (HTPD)
• Creates the ability to react quickly if an “unusual”
observation is made
• All process decisions need to be made keeping large-
scale production in mind

10. 10
MAB PLATFORMS

11. 11
Next generation mAb platforms
• Driver
• High cell culture productivity is increasing interest in ultra-
high loading polishing steps (> 100 mg/mL loading)
Genentech Biogen Millipore proposal
Protein A
Viral Inactivation
Cation-exchange
chromatography
Anion-exchange
chromatography
Viral Filtration
UF/DF
Protein A
Viral Inactivation
AEX flowthrough
No salt Hydrophobic
Interaction
Chromatography
flowthrough
Viral Filtration
UF/DF
Protein A
Viral Inactivation
Anion-exchange
flowthrough
Overloaded cation-
exchange
chromatography
Viral Filtration
UF/DF

12. 12
Which platform should I use?

13. 13
Next generation mAb platforms
• Platform processes for mAbs have hugely facilitated
the growth of mAbs as therapeutic agents
• Rapid clinical entry with lower cost & resource burden
& significant time savings (gene to IND in ~ 9 months)
DS
Process
Platform
DS
Process
Platform
Cell line
diversity
Cell line
diversity
Media/feed
type
diversity
Media/feed
type
diversity
HCP level
variability
HCP level
variability
Cell density
variability
Cell density
variability
HMW level
variability
HMW level
variability
Protein A
Viral Inactivation
AEX Based Polishing
(Flow Through mode)
CEX Based Polishing
Viral Filtration
UF/DF

14. 14
Multimodal chromatography in next
generation mAb platforms
• Mixed-mode has the simultaneous ability to clear HMW and HCP
leading to mAb platforms with wider coverage
• Added advantage of ability to operate over wider conductivity
range for loading
93.0
94.0
95.0
96.0
97.0
98.0
99.0
100.0
0.0 10.0 20.0 30.0 40.0 50.0 60.0 70.0 80.0 90.0 100.0
Capto S ImpAct pH 5.0
Eshmuno CPX pH 5.0
Fractogel SO3 pH 5.0
Capto MMC pH 7.5
Selectivity Curves for HMW Clearance
MainPeak(%)
Increase selectivity
Accumulated Yield (%)
Hydrophobicity scale:
Capto S < Fractogel SO3 < POROS HS50 < Nuvia cPrime < Capto MMC

15. 15
0
50
100
150
200
250
300
350
400
450
500
0.0% 20.0% 40.0% 60.0% 80.0% 100.0%
HCP (ppm)
Recovery
Capto MMC HCP Clearance
25mM Tris pH 7.0 (baseline)
25mM Tris pH 7.0, 5% ethylene glycol
25mM Tris pH 7.0, 50mM arginine
25mM Tris pH 7.0, 50mM NaSCN
25mM Tris pH 7.0, 1M urea
25mM Tris pH 7.0, 1M ammonium sulfate
25mM Tris pH 7.0, 0.1M NaCl
25mM Tris pH 7.0, 0.5M ammonium sulfate
25mM Tris pH 7.0, 0.1M NaCl, 1M urea
Washes that
disrupt
protein-protein
interactions
Conventional washes
log k’ = A – Blog(csalt) + C(csalt) k’ = (tr – tm )/tm
Wolfe, L., Barringer, C., Mostafa, S., Shukla, A.
Multimodal chromatography: characterization of protein binding
and selectivity enhancement through mobile phase modulators,
Journal of Chromatography A, 1340, 151-156, 2014.
Multimodal chromatography

16. 16
Success of mAb platforms that
include multimodal chromatography
• Can successfully accommodate wide range of cell
lines and cell culture feed streams into a single
downstream platform
• Cell lines from KBI, Bioceros, Selexis, Cellca,
Excellgene, Antitope, Life Technologies
0.0%
1.0%
2.0%
3.0%
4.0%
5.0%
6.0%
7.0%
8.0%
ProA AEX CEX BDS
%HMW
mAb Platform HMW Clearance
mAb A
mAb B
mAb C
mAb D
mAb E
mAb F
mAb G
mAb H
mAb I
0
5000
10000
15000
20000
25000
ProA AEX CEX BDS
rHCP(ppm)
mAb Platform rHCPClearance
mAb A
mAb B
mAb C
mAb D
mAb E
mAb F
mAb G
mAb H
mAb I

17. 17
Shukla, A.; Wolfe, L.;
Mostafa, S.; Norman, C.
Evolving trends in mAb
production processes,
Bioengineering and
Translational Medicine,
2(1), 58-69, 2017.

18. 18
High capacity Protein A chromatography resins
Amsphere A3 Protein A chromatography
Resin Vendor Matrix Ligand
Modified Protein A
domain
Mean particle
size (µm)
MabSelect SuReTM GE Healthcare
Highly cross‐linked
agarose
Alkali‐stabilized
rProtein A
B domain 85
MabSelect SuReTM LX GE Healthcare
Highly cross‐linked
agarose
Alkali‐stabilized
rProtein A
B domain 85
ToyopearlTM AF‐rProtein
A HC‐650F† Tosoh Polymethacrylate
Alkali‐stabilized
rProtein A
C domain 45
EshmunoTM A† EMD Millipore
Cross‐linked
Polyvinyl Ether
Alkali‐stabilized
rProtein A
C domain 50
AmsphereTM A3† JSR Life
Sciences
Polymethacrylate
Alkali‐stabilized
rProtein A
C domain 50
1 2 3 4 5 6
0
10
20
30
40
50
60
70
80
mAb2
DBC(g/L)
Residence Time (min)
MabSelect SuRe
MabSelect SuRe LX
rProtein A HC-650F
Eshmuno A
Amsphere A3
mAb1 mAb2 mAb3 mAb4
0
1000
2000
3000
8000
9000
10000
HCPLevel(ppm)
MabSelec SuRe
MabSelect SuRe LX
rProtein A HC-650F
Eshmuno A
Amsphere A3

19. Chromassette® introduction
• Easy to stack
• Easy to store
• Easy to configure
• Open source: packed with all commercial resins
19
A pre-packed stackable supported device that enables
high flow rates with current chromatographic resins

20. Feed Distributor
Eluent
Distributor
Top
Plate
Bottom
Plate
Scaffold
Structural Support of
chrom. bed packed within
void space.
Frit
Frit
Chromassette® introduction

21. 21
Cell therapies
Kite
Novartis
Bluebird
Juno
Gene therapy
Oligonucleotides (Ionis)
RNA (Alnylam, Moderna)
Gene editing CRISPR/ZFP (Sangamo)
DNA (Spark, Bluebird)
Evolution in therapeutic proteins
Conventional mAbs  Next generation mAbs
Monotherapies  Combination therapies
bispecific
Novel fusions
Antibody
fragments
Novel scaffolds

22. 22
Diversity of programs enabled by KBI
• Biotech industry innovation switching from conventional mAbs to
bispecifics and various kinds of fusion proteins
• Need for platforms for non-mAbs has arisen
• KBI has been successful in developing platforms for HIV vaccine
proteins, bispecific antibodies and complex Fc fusion proteins

23. • Accelerated development approaches critical for non‐
platform molecules or where platform needs to be defined
Rameez, S.; Mostafa, S. S.; Miller, C.; Shukla, A. A.
High-throughput miniaturized bioreactors for cell
culture
process development: Reproducibility, scalability
and control.
Biotechnology Progress 2014, (30): 718-727.

24. 24
Shukla, A.A., Rameez,
S., Wolfe, L.S., Oien, N.
High-throughput process
development for
biopharmaceuticals,
Advances in Biochemical
Engineering &
Biotechnology, 2017 (in
press).

25. 25
RAPID PROCESS
CHARACTERIZATION AND
SCALE-DOWN VALIDATION
STUDIES

26. 26
Process Design Space
 Higher level of assurance of
product quality
 Manufacturing Efficiency and
Flexibility
 Continuous process
improvement while maintaining
product quality
Characterization Space
Design space
Control space
Design Space (ICH Q8, 2006): The multidimensional
combination and interaction of input variables (e.g., material
attributes) and process parameters that have been
demonstrated to provide assurance of quality.
Need a high throughput
scale-down model for
the process

27. 27
Accelerating process characterization &
scale-down validation studies
• Small-scale bioreactors (1-10L working volume) have
been the traditional scale-down model in industry till
date
• Accelerating PC/PV studies requires a high-
throughput scale-down model
• Ambr250 as a scale-down model for cell culture
processes

28. 28
Matching key process indicators in SDMs
Comparison of time courses for viable cell growth and lactate profiles for two recombinant CHO cell lines in
ambr™ SDMs for a mAb and a Biosimilar. Matching cell growth and lactate profiles for CHO cell lines
producing a mAb and Biosimilar respectively were key process indicators and in turn dictated the process
yield and product quality.

29. 29
Comparison of SDMs across scales

30. 30
Accelerated Upstream PC Timelines with
high-throughput SDMs
Month 1.5
SDMQ USP
Month 5.5
 N-1/N-2 Screening
(40 x 3L Seed)
Harvest PC Work
12 -15 Harvest conditions
Month 0
 Raw Materials and Worst Case
(20 x 3L and 1 round of ambr250 runs: 24 vessels)
 Main Stage PC
(3 rounds of ambr250 runs: 72 vessels)
 Inoculum Studies
(100 Shake Flasks and 4 Wave Runs)
Worst-case Linkage
USP/DSP
Month 7.0

31. 31
HOW INNOVATION CAN HELP
REDUCE COSTS

32. 32
Gottschalk, U., Brorson,
K., Shukla, A. The need
for innovation in
biomanufacturing, Nature
Biotechnology, 30(6),
489-492, 2012.
Gottschalk, U., Brorson,
K., Shukla, A. Innovation
in biomanufacturing – the
only way forward,
Pharmaceutical
Bioprocessing, 1(2), 141-
157, 2013.

33. 33
COGS for recombinant protein drugs
$/g
$ 1000-
2000/g
e.g.
1980s
to 1990s $ 100-200/g
High titers
Improved cell
lines
Improved media
High capacity
resins
>10,000L scale
Bioreactors &
Single
Use Mfg.
$ 10-20/g
Continuous production
processes?
Rapid cycling chromatography?
Non-chromatographic purification?

34. 34
Let us not miss innovations that
can really make a difference