Access the interactive recording: https://bit.ly/386d4fh
Abstract:
The bioprocessing industry is driving towards intensified processes to reduce cost of goods and/or increase productivity. Perfused seed is one specific intensified upstream process that can provide benefits to mAb production. To better understand these benefits, BioSolve process modeling software was used to perform a holistic cost analysis of several different perfused seed train scenarios. The effect of variables such as production/seed ratio, number of production bioreactors, titer, and production duration were evaluated. Results showed that under certain scenarios, perfused seed train options could deliver lower cost of goods, increase product throughput, or a combination of both.
2. 2
The life science business of
Merck KGaA, Darmstadt, Germany
operates as MilliporeSigma
in the U.S. and Canada.
3. Part II of the BioContinuum™ Seed Train Platform webinar series:
Novel Perfusion Filter and Controller for N-1 Application
Considerations for a scalable cell retention device for perfusion applications
Case study demonstrating capabilities of novel perfusion filter and controller for intensified upstream
processing
Previous Webinar
3
6. Speed
Reduce new facility build
times by 70%. Compress
production lead time by
80%.
Quality
10X robustness.
90% reduction in cost of
poor quality.
Flexibility
Reduce product change-over
time by 90%.
Cost
90% reduction in cost to
manufacture and CAPEX.
Business
Drivers
Market
Growth
Uncertainty
New Product
Classes
Cost
Pressure
Market
Trends
Market Trends, Business Drivers and Key Enablers to
Drive Next Generation BioProcessing:
Process
Intensification
Process
Analytics
Software &
Automation
Key
Enablers
Single Use
6
7. less
footprint
Less
capital
reduction in
construction time
reduction
OPEX
Reduction in
cost of quality
90%
90%
70%
60%
90%
Paradigm Shift:
The Facility of the Future will require a
transformation in how biologics are
developed and manufactured
Process
Format
Analytics
Digital Controlled
Inline
Single Use
Intensified
Integrated
Real Time
Closed
Connected
Autonomous
Predictive
Sterile
Continuous
7
8. Upstream Process Intensification for mAbs
A Multitude of Strategies all based on perfusion technology
Batch
x
1-2ml
each
10-30 x106 vc.ml-1
N-1
or
x
Batch
N
Fed Batch
MCB/MWCB INOCULATION TRAIN
SEED TRAIN
PRODUCTION
CRD
Perfusion
CRD
50-100 x106vc.ml-1
or or
4.5ml 50-500ml bagCRD
CRD
Perfusion
N
CRD
Continuous protein harvest
• Steady State Perfusion
• Dynamic Perfusion
Single protein harvest
• High Seed Fed batch
• Ultrahigh VCD Fed-Batch
• Concentrated Fed-Batch
Production
medium
Cryo Medium
Perfused Seed Train Processes
Can be utilized with any production method
Perfusion-based Production and Harvest processes
Continuous vs. single protein harvest, High cell densities, long durations
Expansion
medium
Cell Culture Media Formulations
Enabling intensified seed and production trains
High Qp
Low CSPR
Low OUR
High Shear resistance
Perfusion Optimized Cell Line
Maximizing protein yield and quality in perfusion operations
8
9. Definition
Perfused Seed Train
14 days
10 days
Conventional
Fed-Batch
(~ 0.2 E6 vc/mL)
High Seed
Fed-Batch
(~ 5 E6 vc/mL)
Conventional
(Batch N-1)
3 days3 days3 days3 days
or
5 days
Compressed
Seed Train
High Seed
(N-1 only)
High Seed
(N-1 & N-2)
Compressed Seed Train: facility utilization and reduced footprint
High-Seed: increased manufacturing capacity, facility utilization
8 days
or
3 days3 days3 days
10 days
or
3 days3 days3 days3 days
5 days8 days
or
3 days3 days3 days
9
11. 11
What is Biosolve®?
• Excel-based process model from BioPharm Services
• Calculates process economics
• Database of industry averages with the option for user-
specific information
Benefits of Process Modeling
• Focus on the “holistic process”
• Side-by-side evaluation of alternatives
• Can run “what if”, sensitivity analyses
• Clear communication of value
Unique Intensified Operation Modules of
Process Modeling
• Perfusion seed train and production options
• Continuous chromatography
• Flow through polishing
• Single-pass TFF
Biosolve®
The Benefits of Process Modeling (Value Modeling)
12. Perfused Seed Train
Model Assumptions
14 days
10 days
Conventional
Fed-Batch
(~ 0.2 E6 vc/mL)
High Seed
Fed-Batch
(~ 5 E6 vc/mL)
Conventional
(Batch N-1)
3 days3 days3 days3 days
or
5 days
Compressed
Seed Train
High Seed
(N-1 only)
High Seed
(N-1 & N-2)
8 days
or
3 days3 days3 days
10 days
or
3 days3 days3 days3 days
5 days8 days
or
3 days3 days3 days
12
Models to be
tested:
13. Perfused Seed Train
Model Assumptions
14 days
10 days
Conventional
Fed-Batch
(~ 0.2 E6 vc/mL)
High Seed
Fed-Batch
(~ 5 E6 vc/mL)
Conventional
(Batch N-1)
3 days
10L
3 days
100mL
3 days
30mL
5 days
60L,200L
Compressed
Seed Train
High Seed
(N-1 only)
High Seed
(N-1 & N-2)
13
Models to be
tested:
3 days
400mL
3 days
2L
125mL 500mL 2X500mL 2X3L 20L
200L
2000L
7 days
10L
20L
3 days
10L
20L
9 days
60L,200L
200L
7 days
10L
20L
4 days
200L
200L
Same as above
Same as above
Same as above
2000L
Each model contains a
traditional downstream!
14. Scope
14
• Constant production titer
• Constant production duration
• Constant production titer, 3:1 P/S
• Constant production titer, 2:1 P/S
• Constant production titer, 1:1 P/S
• Constant production duration, 3:1 P/S
• Constant production duration, 2:1 P/S
• Constant production duration, 1:1 P/S
Proposed Scenarios
Models: Conventional (Batch N-1), Compressed Seed Train, High Seed (pN-1), High Seed (pN-2/pN-1)
Goal Gain an understanding of the potential benefits of perfused seed trains through Biosolve®
Single Bioreactor Multiple Bioreactors (6)
14 days
4 g/L
vs. 10 days
4 g/L
Traditional High Seed
14 days
>4 g/L
15. Scope
15
• Constant production titer, 3:1 P/S
• Constant production titer, 2:1 P/S
• Constant production titer, 1:1 P/S
• Constant production duration, 3:1 P/S
• Constant production duration, 2:1 P/S
• Constant production duration, 1:1 P/S
Proposed Scenarios
Models: Conventional (Batch N-1), Compressed Seed Train, High Seed (pN-1), High Seed (pN-2/pN-1)
Goal Gain an understanding of the potential benefits of perfused seed trains through Biosolve®
Multiple Bioreactors (6)
6 Bioreactors2 Seed Trains
3:1 Ratio
16. Scope
16
• Constant production titer, 3:1 P/S
• Constant production titer, 2:1 P/S
• Constant production titer, 1:1 P/S
• Constant production duration, 3:1 P/S
• Constant production duration, 2:1 P/S
• Constant production duration, 1:1 P/S
Proposed Scenarios
Models: Conventional (Batch N-1), Compressed Seed Train, High Seed (pN-1), High Seed (pN-2/pN-1)
Goal Gain an understanding of the potential benefits of perfused seed trains through Biosolve®
Multiple Bioreactors (6)
6 Bioreactors3 Seed Trains
2:1 Ratio
17. Scope
17
• Constant production titer, 3:1 P/S
• Constant production titer, 2:1 P/S
• Constant production titer, 1:1 P/S
• Constant production duration, 3:1 P/S
• Constant production duration, 2:1 P/S
• Constant production duration, 1:1 P/S
Proposed Scenarios
Models: Conventional (Batch N-1), Compressed Seed Train, High Seed (pN-1), High Seed (pN-2/pN-1)
Goal Gain an understanding of the potential benefits of perfused seed trains through Biosolve®
Multiple Bioreactors (6)
6 Bioreactors6 Seed Trains
1:1 Ratio
19. Single Bioreactor, Constant Production Titer
19
CoG’s ($/g) 116.39 116.80 101.17 101.64
Throughput
(kg/yr)
90.18 90.18 125.25 125.25
Batches/yr 18 18 25 25
Assumptions
• 1 X 2000L Bioreactor
• 1 X Seed train
• 4 g/L titer
• Traditional: 14 day production
• High Seed: 10 day production
• ~13.5% reduction in CoG’s by
switching to perfused seed train
• Increased throughput as well;
decreased production duration
increases batches/yr
• Cost reduction driven by higher
throughput; brings down capital
High Seed Fed Batch
Traditional Fed Batch
20. Single Bioreactor, Constant Production Duration
20
Assumptions
• 1 X 2000L Bioreactor
• 1 X Seed train
• Traditional: 14 day production
• High Seed: 14 day production
• Experimental data suggests titer can
increase 2-3X through high seeding
production
• Traditional seed train was compared
to perfused N-1 seed train with
varying production titers
• Higher throughput drives down all
costs; increased product mass per
batch
High Seed Fed Batch
CoG’s ($/g)
116.4 101.2 120.1 90.6 75.6 64.0 57.0
Throughput
(kg/yr)
90.18 125.25 90.18 135.27 180.36 225.45 270.54
Batches/yr
18 25 18 18 18 18 18
21. Single Bioreactor, Constant Production Duration
21
Assumptions
• 1 X 2000L Bioreactor
• 1 X Seed train
• Traditional: 14 day production
• High Seed: 14 day production
Seed Train Titer
Total
CoG’s
($/g)
%
Reduction
Traditional 4 g/L 116.39
pN-1 6 g/L 90.56 22%
pN-1 8 g/L 75.56 35%
pN-1 10 g/L 63.98 45%
pN-1 12 g/L 56.99 51%
High Seed Fed Batch
CoG’s ($/g)
116.4 101.2 120.1 90.6 75.6 64.0 57.0
Throughput
(kg/yr)
90.18 125.25 90.18 135.27 180.36 225.45 270.54
Batches/yr
18 25 18 18 18 18 18
22. Single Bioreactor Scenarios - Summary
22
Constant Production Titer
14 days
4 g/L
vs. 10 days
4 g/L
❖ High Seed options increase batch/yr
and throughput
18 –> 25 batch/yr
90 –> 125 kg/yr
❖ Drives capital CoG’s down, total CoG’s
down 13.5%
Constant Production Duration
Traditional High Seed
❖ Greater benefit observed through
keeping constant production duration
❖ Overall CoG’s reduced 22-51%
❖ Throughput increase 50-200%;
proportional to titer increase
❖ No change in batches/yr
Conventional
(Batch N-1)
Compressed
Seed Train
High Seed
(N-1 only)
High Seed
(N-1 & N-2)
14 days
4 g/L
vs. 14 days
>4 g/L
Traditional High Seed
Conventional
(Batch N-1)
Compressed
Seed Train
High Seed
(N-1 only)
High Seed
(N-1 & N-2)
24. 3:1 Production/Seed Train Ratio
24
Assumptions
• 6 X 2000L Bioreactor
• 2 X Seed train
• 4 g/L titer
• Traditional: 14 day production
• High Seed: 10 day production
CoG’s ($/g) 67.38 77.25 85.46 82.80
Throughput
(kg/yr) 495.99 315.63 275.55 315.63
Batches/yr 99 63 55 63
• No cost or throughput
benefit for 3:1
production/seed train ratio
• With 6 bioreactors,
production is no longer
bottleneck
25. 3:1 Production/Seed Train Ratio
25
Assumptions
• 6 X 2000L Bioreactor
• 2 X Seed train
• 4 g/L titer
• Traditional: 14 day production
• High Seed: 10 day production
• No cost or throughput
benefit for 3:1
production/seed train ratio
• With 6 bioreactors,
production is no longer
bottleneck
Process
Bottleneck
Unit Op
Bottleneck
Time
(days/batch)
Batches/yr
Traditional N-1 2.64 99
Compressed Seed Fed
Batch
Perfused
Rocker
4.12 63
Perfused Seed (pN-1) N-1 4.76 55
Perfused Seed (pN-2/pN-1) N-2 Rocker 4.12 63
26. 2:1 Production/Seed Train Ratio
26
Assumptions
• 6 X 2000L Bioreactor
• 3 X Seed train
• 4 g/L titer
• Traditional: 14 day production
• High Seed: 10 day production
CoG’s ($/g) 65.88 67.58 75.03 73.08
Throughput
(kg/yr) 551.11 475.95 410.82 475.95
Batches/yr 110 95 82 95
• Similarly to 3:1, no cost or
throughput benefit for 2:1
production/seed train ratio
• Bottlenecking effect is less
pronounced, due to extra
seed train
27. 2:1 Production/Seed Train Ratio
27
Assumptions
• 6 X 2000L Bioreactor
• 3 X Seed train
• 4 g/L titer
• Traditional: 14 day production
• High Seed: 10 day production
Process
Bottleneck
Unit Op
Bottleneck
Time
(days/batch)
Batches/yr
Traditional Production 2.39 110
Compressed Seed Fed
Batch
Perfused
Rocker
2.74 95
Perfused Seed (pN-1) N-1 3.17 82
Perfused Seed (pN-2/pN-1) N-2 Rocker 2.74 95
• Similarly to 3:1, no cost or
throughput benefit for 2:1
production/seed train ratio
• Bottlenecking effect is less
pronounced, due to extra
seed train
28. 1:1 Production/Seed Train Ratio
28
Assumptions
• 6 X 2000L Bioreactor
• 6 X Seed train
• 4 g/L titer
• Traditional: 14 day production
• High Seed: 10 day production
CoG’s ($/g)
66.88 65.35 65.06 66.52
Throughput
(kg/yr)
551.11 546.10 761.53 761.53
Batches/yr
110 109 152 152
• Negligible CoG’s differences,
though significant increase in
throughput for high seed fed
batch
• With 1:1 ratio, production
becomes bottleneck again;
decreased HiSeed production
duration increases throughput
29. 1:1 Production/Seed Train Ratio
29
Assumptions
• 6 X 2000L Bioreactor
• 6 X Seed train
• 4 g/L titer
• Traditional: 14 day production
• High Seed: 10 day production
Process
Bottleneck
Unit Op
Bottleneck
Time
(days/batch)
Batches/yr
Traditional Production 2.39 110
Compressed Seed Fed
Batch
Production 2.40 109
Perfused Seed (pN-1) Production 1.73 152
Perfused Seed (pN-2/pN-1) Production 1.73 152
• Negligible CoG’s differences,
though significant increase in
throughput for high seed fed
batch
• With 1:1 ratio, production
becomes bottleneck again;
decreased HiSeed production
duration increases throughput
30. Increased Titer with High Seed Production – 3:1 P/S ratio
30
Assumptions
• 3:1 P/S ratio
• 6 X 2000L Bioreactor
• 2 X Seed train
• Traditional: 14 day production
• High Seed: 14 day production
CoG’s ($/g) 67.38 85.46 85.46 62.49 52.01 43.78 39.39
Throughput
(kg/yr) 495.99 275.55 275.55 413.33 551.11 688.88 826.66
Batches/yr 99 55 55 55 55 55 55
High Seed Fed Batch
• As seen earlier, process
bottlenecks reduce batches/yr
for perfused seed options at 3:1
P/S ratio
• Increased titer offsets that
reduction in most cases,
ultimately increasing throughput
• Higher throughput and product
per batch drives down CoG’s
31. Increased Titer with High Seed Production – 2:1 P/S Ratio
31
Assumptions
• 2:1 P/S ratio
• 6 X 2000L Bioreactor
• 3 X Seed train
• Traditional: 14 day production
• High Seed: 14 day production
CoG’s ($/g) 65.88 75.03 75.03 54.67 45.75 38.49 34.64
Throughput
(kg/yr) 551.11 410.82 410.82 616.24 821.65 1027.06 1232.57
Batches/yr 110 82 82 82 82 82 82
High Seed Fed Batch
• Similar trend observed
• With bottleneck effect
diminishing, throughput
increase is larger
32. Increased Titer with High Seed Production – 1:1 P/S Ratio
32
Assumptions
• 1:1 P/S ratio
• 6 X 2000L Bioreactor
• 6 X Seed train
• Traditional: 14 day production
• High Seed: 14 day production
CoG’s ($/g) 66.88 65.06 70.79 51.46 43.08 36.17 32.67
Throughput
(kg/yr) 551.11 761.53 546.10 819.14 1092.19 1365.24 1638.29
Batches/yr 110 152 109 109 109 109 109
High Seed Fed Batch
• Again, for 1:1 P/S ratio,
production becomes
bottleneck
• Batches/yr more or less
constant
• Most significant increase
in throughput
33. Increased Titer with High Seed – Summary (6 Bioreactors)
33
• When considering the potential for increased
titer through high seed fed batch, virtually all
scenarios offer benefits
• For both CoG’s and throughput, most benefit is found
at 1:1 production/seed train ratio
• Decrease CoG’s 23-51%
• Increase throughput 65-230%
% CoG’s Reduction and % Throughout Increase calculated by comparing to Traditional 4 g/L process
34. Multiple Bioreactor Scenarios - Summary
34
Constant Production Titer
14 days
4 g/L
vs. 10 days
4 g/L
❖ No benefit observed in 3:1 and 2:1
P/S scenarios; bottleneck issues
❖ At 1:1 P/S ratio, throughput increase
due to increase batches/yr
109->152 batches/yr
Constant Production Duration
Traditional High Seed
❖ Overall CoG’s decrease and
throughput increase at all P/S ratios
❖ For 3:1 and 2:1 P/S ratios, batches/yr
is reduced, even with CoG’s and
throughput benefits
Conventional
(Batch N-1)
Compressed
Seed Train
High Seed
(N-1 only)
High Seed
(N-1 & N-2)
14 days
4 g/L
vs. 14 days
>4 g/L
Traditional High Seed
Conventional
(Batch N-1)
Compressed
Seed Train
High Seed
(N-1 only)
High Seed
(N-1 & N-2)
35. 35
Implementing perfused seed trains can
bring down cost of manufacturing and
increase product throughput, all while
keeping production bioreactor in fed
batch mode
Perfused seed trains allow for high
seeding of production bioreactor, which
has the potential for achieving higher
titers, especially when incorporating a
strong media platform
Summary
If higher titers observed:
• Adding additional seed trains and production bioreactors at
1:1 ratio brings the most benefit
• CoG’s reduced 24-52%, kg/yr increase 65-230%
• Benefits even at 3:1 and 2:1, with additional benefit of
needing less batches per year
• 3:1 – CoG’s reduced 7-42%, kg/yr increase 11-67%
• 2:1 - CoG’s reduced 19-49%, kg/yr increase 24-149%
If higher titers are not observed:
• Single bioreactor scenarios still see CoG’s reduced by 13.5%
and throughput increase of 39% at shorter high seed FB
duration
• Adding additional seed trains and production bioreactors at
1:1 ratio increases throughput