Suzanne Farid PhD CEng FIChemE

Reader (Associate Professor)

Co-Director EPSRC Centre for Innovative Manufacturing
UCL Biochemical Engineering
s.farid@ucl.ac.uk

ECI Integrated Continuous Biomanufacturing, Barcelona, Spain, 20-24 October 2013

UCL Decisional Tools Research

Operational & Economic Evaluation of
Integrated Continuous Biomanufacturing
Strategies for Clinical & Commercial
mAb Production
2
Engineering Doctorate Project:
Evaluating The Potential of Continuous Processes for Monoclonal
Antibodies: Economic, Environmental and Operational Feasibility

UCL-Pfizer Collaboration (2008-2013)







UCL academic collaborators included: Daniel Bracewell
(ex-)Pfizer collaborators included: Glen Bolton, Jon Coffman

Funding: UK EPSRC, Pfizer







Acknowledgements
James Pollock
UCL
Suzanne Farid
UCL
Sa Ho
Pfizer
3
Decisions Portfolio selection? Process design? Capacity Sourcing? Build single / multi-product facility?
Uncertainties Clinical (e.g. doses, transition probabilities) Technical (e.g. titres, equipment failure) Commercial (e.g. sales forecasts)
Constraints Time Capacity Budget Regulatory Skilled labour
Metrics Speed Ease of scale-up Cost of goods Fit to facility Robustness
Bioprocess Decisional Tools Domain
Biotech Drug Development Cycle
Farid, 2012, In Biopharmaceutical Production Technology, pp717-74
4
Scope of UCL Decisional Tools
Typical questions addressed:
Process synthesis & facility design
Which manufacturing strategy is the most cost-effective?
How do the rankings of manufacturing strategies change with scale?
Or from clinical to commercial production?
Key economic drivers? Economies of scale?
Probability of failing to meet cost/demand targets? Robustness?

Portfolio management & capacity planning
Portfolio selection - Which candidate therapies to select?
Capacity sourcing - In-house v CMO production?
Impact of company size and phase transition probabilities on choices?
5
Systems approach to valuing biotech / cell therapy investment opportunities
Process synthesis and facility design
Capacity planning
Portfolio management
Challenges:
Capturing process robustness under uncertainty & reconciling conflicting outputs
Fed-batch versus perfusion systems (Lim et al, 2005 & 2006; Pollock et al, 2013a)
Continuous chromatography (Pollock et al, 2013b)
Integrated continuous processing (Pollock et al, submitted)
Stainless steel versus single-use facilities (Farid et al, 2001, 2005a &b)
Facility limits at high titres (Stonier et al, 2009, 2012)
Single-use components for allogeneic cell therapies (Simaria et al, 2013)
Adopting efficient methods to search large decision spaces
Portfolio management & capacity planning (Rajapakse et al, 2006; George & Farid, 2008a,b)
Multi-site long term production planning (Lakhdar et al, 2007; Siganporia et al, 2012)
Chromatography sequence and sizing optimisation in multiproduct facilities (Simaria et al, 2012;
Allmendinger et al, 2012)
Integrating stochastic simulation with advanced multivariate analysis
Prediction of suboptimal facility fit upon tech transfer (Stonier et al, 2013; Yang et al, 2013)
Creating suitable data visualization methods
For each of above examples
Scope of UCL Decisional Tools
Farid, 2012, In Biopharmaceutical Production Technology, pp717-74
6
Systems approach to valuing biotech / cell therapy investment opportunities
Process synthesis and facility design
Capacity planning
Portfolio management
Challenges:
Capturing process robustness under uncertainty & reconciling conflicting outputs
Fed-batch versus perfusion systems (Pollock et al, 2013a)
Continuous chromatography (Pollock et al, 2013b)
Integrated continuous processing (Pollock et al, submitted)
Stainless steel versus single-use facilities (Farid et al, 2001, 2005a &b)
Facility limits at high titres (Stonier et al, 2009, 2012)
Single-use components for allogeneic cell therapies (Simaria et al, submitted)
Adopting efficient methods to search large decision spaces
Portfolio management & capacity planning (Rajapakse et al, 2006; George & Farid, 2008a,b)
Multi-site long term production planning (Lakhdar et al, 2007; Siganporia et al, 2012)
Chromatography sequence and sizing optimisation in multiproduct facilities (Simaria et al, 2012)
Integrating stochastic simulation with advanced multivariate analysis
Prediction of suboptimal facility fit upon tech transfer (Stonier et al, 2013; Yang et al, 2013)
Creating suitable data visualization methods
For each of above examples
Scope of UCL Decisional Tools
Farid, 2012, In Biopharmaceutical Production Technology, pp717-74
7
Systems approach to valuing biotech / cell therapy investment opportunities
Process synthesis and facility design
Capacity planning
Portfolio management
Challenges:
Capturing process robustness under uncertainty & reconciling conflicting outputs
Scope of UCL Decisional Tools
Fed-batch versus perfusion systems (Pollock et al, 2013a)
Scenario: New build for commercial mAb prodn
Impact of scale on cost
Impact of titre variability and failures rates on robustness
Continuous chromatography (Pollock et al, 2013b)
Scenario: Retrofit for clinical / commercial mAb prodn
Impact of scale and development phase on cost
Retrofit costs across development phases
Integrated continuous processing (Pollock et al, submitted)
Scenario: New build for clinical / commercial mAb prodn
Impact of hybrid batch/continuous USP and DSP combinations
Impact of development phase, company size and portfolio size

8
Fed-batch versus perfusion culture (New build)

Pollock, Ho & Farid, 2013, Biotech Bioeng, 110(1): 206219
Fed-batch versus perfusion systems (Pollock et al, 2013a)
Scenario: New build for commercial mAb prodn
Impact of scale on cost
Impact of titre variability and failures rates on robustness
9
Fed-batch versus perfusion culture (New build)

Pollock, Ho & Farid, 2013, Biotech Bioeng, 110(1): 206219
Commercial products using perfusion cell culture technologies
10 Pollock, Ho & Farid, 2013, Biotech Bioeng, 110(1): 206219
LEVEL
CONTROL
OFF
ON
AIR
INLET
EXHAUST
ADDITION
PUMP
FLUID
INLET
VALVE
QUICK CONNECT
FILTRATE PUMP
FILTRATE
0.2 MICRON HOLLOW FIBRE FILTER CASSETTE
HOUSING
CONTROLLER
PROCESS VESSEL
DIAPHRAGM
ATF
PUMP
STAND
FILTER
LIQUID
LEVEL
LEVEL
CONTROL
LEVEL
CONTROL
OFF
ON
OFF
ON
AIR
INLET
EXHAUST
ADDITION
PUMP
FLUID
INLET
VALVE
QUICK CONNECT
FILTRATE PUMP
FILTRATE
0.2 MICRON HOLLOW FIBRE FILTER CASSETTE
HOUSING
CONTROLLER
PROCESS VESSEL
DIAPHRAGM
ATF
PUMP
STAND
FILTER
LIQUID
LEVEL
LEVEL
CONTROL
OFF
ON
AIR
INLET
EXHAUST
ADDITION
PUMP
FLUID
INLET
VALVE
QUICK CONNECT
FILTRATE PUMP
FILTRATE
0.2 MICRON HOLLOW FIBRE FILTER CASSETTE
HOUSING
CONTROLLER
PROCESS VESSEL
DIAPHRAGM
ATF
PUMP
STAND
FILTER
LIQUID
LEVEL
LEVEL
CONTROL
LEVEL
CONTROL
OFF
ON
OFF
ON
AIR
INLET
EXHAUST
ADDITION
PUMP
FLUID
INLET
VALVE
QUICK CONNECT
FILTRATE PUMP
FILTRATE
0.2 MICRON HOLLOW FIBRE FILTER CASSETTE
HOUSING
CONTROLLER
PROCESS VESSEL
DIAPHRAGM
ATF
PUMP
STAND
FILTER
LIQUID
LEVEL
SPIN
FILTER
LIQUID
LEVEL
Spin-filter Perfusion
PRO:
CON:
Investment
DSP consumable cost
Equipment failure rate
USP consumable cost
Scale limitations
Validation burden
Compare the cost-effectiveness and robustness of fed-batch and perfusion cell
culture strategies across a range of titres and production scales for new build
ATF Perfusion
Steady state cell densities
Failure rates
LEVEL
CONTROL
OFF
ON
AIR
INLET
EXHAUST
ADDITION
PUMP
FLUID
INLET
VALVE
QUICK CONNECT
FILTRATE PUMP
FILTRATE
0.2 MICRON HOLLOW FIBRE FILTER CASSETTE
HOUSING
CONTROLLER
PROCESS VESSEL
DIAPHRAGM
ATF
PUMP
STAND
FILTER
LIQUID
LEVEL
LEVEL
CONTROL
LEVEL
CONTROL
OFF
ON
OFF
ON
AIR
INLET
EXHAUST
ADDITION
PUMP
FLUID
INLET
VALVE
QUICK CONNECT
FILTRATE PUMP
FILTRATE
0.2 MICRON HOLLOW FIBRE FILTER CASSETTE
HOUSING
CONTROLLER
PROCESS VESSEL
DIAPHRAGM
ATF
PUMP
STAND
FILTER
LIQUID
LEVEL
Fed-batch versus perfusion culture (New build)
Scenario trade-offs: FB v SPIN v ATF
11
Cell
Culture
Suite
DSP
Suite
Viral
Secure
Suite
Seed #1
Seed #2
CC
Cent
DF
UF
ProA
VI
CEX
UFDF
VRF
AEX
UFDF
Seed #1
Seed #2
CC
DF
Seed #1
Seed #2
CC
ProA
VI
CEX
UFDF
VRF
AEX
UFDF
Pool
ProA
VI
CEX
UFDF
VRF
AEX
UFDF
Pool
Suites FB SPIN ATF
Variable FB SPIN ATF
Reactor type SS/SUB SS SUB
Cell culture time (days) 12 60 60
Max VCD (10
6
cells/ml) 10 15 50
Max bioreactor vol. (L) 20,000 2000 1500
Max perf. rate (vv/day) 1 1.5
Process yield 65% 68% 69%
Annual # batches 22 5 5
Product conc. (g/L) 2 10 20% FB 45% FB
Productivity (mg/L/day) 170-850 2 x FB 6.5 x FB
Pollock, Ho & Farid, 2013, Biotech Bioeng, 110(1): 206219
Fed-batch versus perfusion culture (New build)
Key assumptions
12
Comparison of the cost of goods per gram for an equivalent fed-batch titre of 5 g/L
Pollock, Ho & Farid, 2013, Biotech Bioeng, 110(1): 206219
Fed-batch versus perfusion culture (New build)
Results: Impact of scale on COG
= Indirect
= Material
= Labour
Critical cell density difference for ATF to compete with FB - x3 fold.
13
Process event p(Failure) Consequence
Fed-batch culture contamination 1 % Batch loss
Spin-filter culture contamination 6 %
Batch loss & discard two
pooled perfusate volumes
Spin-filter filter failure 4 %
Batch loss & no pooled
volumes are discarded
ATF culture contamination 6 %
Batch loss & discard two
pooled perfusate volumes
ATF filter failure 2 %
Replace filter & discard next 24
hours of perfusate
In process filtration failure general 5 % 4 hour delay & 2% yield loss
In process filtration failure post viral inactivation 20 % 4 hour delay & 2% yield loss
Pollock, Ho & Farid, 2013, Biotech Bioeng, 110(1): 206219
Fed-batch versus perfusion culture (New build)
Uncertainties and failure rates
14
Annual throughput and COG distributions under uncertainty
500kg demand, 5g/L titre
Pollock, Ho & Farid, 2013, Biotech Bioeng, 110(1): 206219
Fed-batch versus perfusion culture (New build)
Results: Impact of variability on robustness
15
Annual throughput and COG distributions under uncertainty
500kg demand, 5g/L titre
Pollock, Ho & Farid, 2013, Biotech Bioeng, 110(1): 206219
Fed-batch versus perfusion culture (New build)
Results: Impact of variability on robustness
16
1. FB = ATF
2. SPIN

1. ATF
2. FB
3. SPIN
1. FB
2. ATF
3. SPIN
Economic
benefits
dominate
Operational
benefits
dominate
Pollock, Ho & Farid, 2013, Biotech Bioeng, 110(1): 206219
Fed-batch versus perfusion culture (New build)
Results: Reconciling operational and economic benefits
fed-batch, -- spin-filter, ATF
17
Continuous chrom: clinical & commercial (Retrofit)

Pollock, Bolton, Coffman, Ho, Bracewell, Farid, 2013, J Chrom A, 1284: 17-27
Continuous chromatography (Pollock et al, 2013b)
Scenario: Retrofit for clinical / commercial mAb prodn
Impact of scale and development phase on cost
Retrofit costs across development phases
18
Technology Evaluation
18
Load
FT
Wash Load
FT
1 ml scale-down
evaluation
3C-PCC system
validation
Discrete event
simulation tool
0%
20%
40%
60%
80%
100%
0 50 100 150
m
A
b

B
r
e
a
k
t
h
r
o
u
g
h

Challenge Load (mg/ml)
100 cm/hr (14.3 mins)
230 cm/hr (6.6 mins)
300 cm/hr (5 mins)
500 cm/hr (3 mins)
Mass balance,
scale-up &
scheduling
equations
Continuous chrom: clinical & commercial (Retrofit)

19
19
3C-PCC
CV = 3 x 1 mL
Titre = 2 g/L
t
res
= 6.6 mins
t
Switch
= 200 mins
t
rampup
= 330 mins
t
rampdown
= 300 mins
ramp-up ramp-down Switch time
Continuous chrom: clinical & commercial (Retrofit)
Example Chromatogram
20
Acidic Designated Basic
Cycle (100 cycles)
19.3 % 75.0 % 5.7 %
Batch (3 cycles)
18.4 % 74.7 % 6.8 %
3C-PCC (6 runs)
18.3 % 75.8 % 5.9 %
HMW Designated LMW
Cycle (100 cycles)
0.7 % 97.6 % 1.7 %
Batch (3 Cycles)
1.0 % 96.9 % 2.1 %
3C-PCC (6 runs)
0.4 % 98.0 % 1.6 %
CEX - HPLC
SEC - HPLC
20
Continuous chrom: clinical & commercial (Retrofit)
Product Quality (Elution peak)
21
Technology Evaluation
21
Load
FT
Wash Load
FT
1 ml scale-down
evaluation
3C-PCC system
validation
Discrete event
simulation tool
0%
20%
40%
60%
80%
100%
0 50 100 150
m
A
b

B
r
e
a
k
t
h
r
o
u
g
h

Challenge Load (mg/ml)
100 cm/hr (14.3 mins)
230 cm/hr (6.6 mins)
300 cm/hr (5 mins)
500 cm/hr (3 mins)
Mass balance,
scale-up &
scheduling
equations
Continuous chrom: clinical & commercial (Retrofit)

22
Day 1 Day 2 Day 3 Day 4 Day 5 Day 6
PA
(1 cycle)
PA
(2 cycle)
PA
(2 cycle)
AEX VRF UFDF
Proof-of-concept (Phase I & II) ~ 4kg DS for the average mAb
1,2
1800L (wv) Fed-batch @ 2.5g/L
Protein A resin costs
~ 60% Direct manufacturing costs
~ $250k per molecule
1. Simaria, Turner & Farid, 2012, Biochem Eng J, 69, 144-154
2. Bernstein, D. F.; Hamrell, M. R. Drug Inf. J. 2000, 34, 909917.
Continuous chrom: clinical & commercial (Retrofit)
Early phase DS manufacture challenges
Pollock, Bolton, Coffman, Ho, Bracewell, Farid, 2013, J Chrom A, 1284: 17-27
23
Standard 3C-PCC
5 cycles 17 cycles
31.4L 3 x 4.9L = 14.7L
$ 250K resin $ 118K resin
53% reduction in resin volume
40% reduction in buffer volume
x2.3 increase in man-hours

Load Wash
Load
Proof-of-concept (Phase I & II) ~ 4kg DS for the average mAb (2.5g/L)
24 hour shift
8 hour shift
Continuous chrom: clinical & commercial (Retrofit)
Results: Economic Impact Protein A
24
PA costs
Other Costs
1 x 4kg 4 x 10kg 20 x 10kg
Pollock, Bolton, Coffman, Ho, Bracewell, Farid, 2013, J Chrom A, 1284: 17-27
Continuous chrom: clinical & commercial (Retrofit)
Results: Impact of scale on direct costs
25
PoC
(1 x 4kg)
PIII &
Commercial
(4 x 10kg)
STD: KTA process (15-600L/hr)
+ 0.4m column
4C-PCC (15-600L/hr)
+ 4 x 0.2m columns
STD: KTA process (45-1800L/hr)
+ 0.5m column
4C-PCC (15-600L/hr)
+ 4 x 0.3m columns
x3.3 Investment

~25 PIII batches
or
~ 8 PoC batches
x4 Investment

~8 PoC batches
Continuous chrom: clinical & commercial (Retrofit)
Results: Impact of development phase on retrofitting investment
26
Integrated continuous processes (New build)
Scenarios: Alternative integrated USP and DSP flowsheets
DSP scheduling

a) batch process
sequence


b) continuous +
batch process
sequence


c) continuous
process
sequence
Pollock, Ho & Farid, submitted
Integrated continuous processing (Pollock et al, submitted)
Scenario: New build for clinical / commercial mAb prodn
Impact of hybrid batch/continuous USP and DSP combinations
Impact of development phase, company size and portfolio size

27
Integrated continuous processes (New build)
Results: Impact of development phase and company size on optimal
Strategies USP Capture Polishing
Base case Fed-batch Batch Batch
FB-CB Fed-batch Continuous Batch
ATF-CB ATF perfusion Continuous Batch
FB-CC Fed-batch Continuous Continuous
ATF-CC ATF perfusion Continuous Continuous
Continuous
USP
+
Continuous
Capture
+
Continuous
Polishing
Batch
USP
+
Continuous
Capture
+
Batch
Polishing
28
Summary
Process economics case study insights:
Fed-batch versus perfusion culture for new build
Economic competitiveness of perfusion depends on cell
density increase achievable and failure rate
Continuous chromatography retrofit
Continuous capture can offer more significant savings
in early-stage clinical manufacture than late-stage
Integrated continuous processes for new build
Integrated continuous processes offer savings for
smaller portfolio sizes and early phase processes
Hybrid processes (Batch USP, Continuous Chrom) can
be more economical for larger / late phase portfolios
Suzanne Farid PhD CEng FIChemE
Reader (Associate Professor)
Co-Director EPSRC Centre for Innovative Manufacturing
UCL Biochemical Engineering
s.farid@ucl.ac.uk

ECI Integrated Continuous Biomanufacturing, Barcelona, Spain, 20-24 October 2013

UCL Decisional Tools Research

Operational & Economic Evaluation of
Integrated Continuous Biomanufacturing
Strategies for Clinical & Commercial
mAb Production
31
Backup

32
3 Column Periodic Counter Current Chromatography
Load
FT
Wash/
Elution
Load
FT
Load FT Wash/
Elution
Pollock, Bolton, Coffman, Ho, Bracewell, Farid, 2013, J Chrom A, 1284: 17-27
Continuous chrom: clinical & commercial (Retrofit)
33
Load
FT
Load FT Wash
40 g/L
65 g/L
FT
Load FT Wash/
Elution
Load
Wash/
Elution
Pollock, Bolton, Coffman, Ho, Bracewell, Farid, 2013, J Chrom A, 1284: 17-27
3 Column Periodic Counter Current Chromatography
Continuous chrom: clinical & commercial (Retrofit)
34
-40%
e-factor
(kg/ kg of protein)
STD 3C-PCC Difference
Water 5900 5250 -11%
Consumable 24.5 13.7 -44%
Proof-of-concept (Phase I & II) ~ 4kg DS for the average mAb (2.5g/L)
STD
3C-PCC
Pollock, Bolton, Coffman, Ho, Bracewell, Farid, 2013, J Chrom A, 1284: 17-27
Continuous chrom: clinical & commercial (Retrofit)
Results: Environmental Impact
35
Large
FB + Cont
Chrom
FB + Cont
Chrom
FB + Cont
Chrom
FB + Cont
Chrom
Medium
ATF + Cont
Chrom
ATF + Cont
Chrom
ATF + Cont
Chrom
FB + Cont
Chrom
Small
ATF + Cont
Chrom
ATF + Cont
Chrom
ATF + Cont
Chrom
FB + Cont
Chrom
Pre-clinical PoC PIII Commercial
C
o
m
p
a
n
y

S
i
z
e
Manuf acturing Scale
Integrated continuous processes (New build)
Results: Impact of development phase and company size on optimal
Strategies USP Capture
Base case Fed-batch Batch
FB-CB Fed-batch Continuous
ATF-CB ATF perfusion Continuous
FB-CC Fed-batch Continuous
ATF-CC ATF perfusion Continuous
Continuous
USP
+
Continuous
Capture
Batch
USP
+
Continuous
Capture
36
Impact of Resin Life Span
(MabSelect x100 cycles)
Standard cycling study (40mg/ml)

Column regeneration (NaOH)

100% breakthrough cycling study
x2.2 the load volume vs. standard
36
19% loss in capacity
12% loss in capacity
30% loss in capacity
Insignificant loss < 15 cycles
37
Commercial Manufacture Feasibility (3C-PCC
@ 5g/L)
37
Batch 11 surpasses harvest hold
time
Batch 6 surpasses pool vessel
volume
Increasing cycle number Increasing cycle number
16 38 22 16 38 19