Viable Cell Density(M cells/mL)

Simplify Upstream Process Intensification From PD to Manufacturing

Higher Productivity

2-3×> 30%

Lower COGS

Source: Data derived from Biosolve modelling

High-Density
Cryobag

Downstream Processing
CRD--

10050Ambr® n=45L n=2

100

30
30
2020
10
10510152030

25Time calculated (days)

97.595Ambr
n=2Ambr®®(N=2)
55L
L n=1
(N=1)

92.5
90

000555A
B
B

101010151515202020

Time calculated (days)
TimecalculatedTime(days)
Time calculated
calculated (days)(days)

252525510959520®®
Ambr
n=2AmbrAmbr®(N=2)
n=255LL(N=1)
n=15L n=1

15202530

Time calculated (days)
CC
150150Coagulation factor, 30-day process, ATF mode,parallel 5 L runs
100
100Automated bleeding to match reference process VCD
Equivalent performance to 5 L bioreactor for
5050A) VCD, B) Viability and C) normalized product titerAmbr®
® n=4Ambr
n=455C

1010

C 150

40
97.5
97.5
30

1515Time
Time calculated
calculated (days)(days)

20205L n=2n=25L
2525300310050Ambr®® n=4(N=4)
Ambr

55L
L (N=2)n=201010
101515
15Timecalculated(days)Time
calculated(days)Time
calculated(days)Time
calculated(days)

B

00030 00

3030

50
10010055
50

2020
20030
30
32525
250

510

15202530

Time(days)
Timecalculated
calculated (days)

100

--

6. Flexible Seed Train Options

N-stage

50 L Cell Bag
(N-2)

500 L

505000

B0B

4040

®

090
90 000Requirements include robust cell line and media suitable for long culture durations & high VCDs of perfusion, scalable
performance from clone screening to manufacturing along with recipes & PAT sensors for process control
N-1 Perfusion

1092.592.5102. Upstream Process Intensification Strategies

Fully closed seed-train

20

Bioreactor
Bioreactortitre
titre

Benefits of Process Intensification:
Enables faster drug development
Increases the efficiency and productivity in GMP manufacturing
Applicable to any process irrespective of moleculeCan be step-wise (per unit operation) or end-to-end for maximum impact
Other terms used in the same context: “continuous”, “connected” and“integrated”

50 - 70%

Flexible Smaller Footprint

30AA

Cedex Viability (%)----< 2 yearsA

Process intensification is a holistic framework to maximize
overall productivity of the unit operation(s), the manufacturing
process and | or the facility output for biomanufacturing.

Cedex Viability (%)

Faster Buildout Time405. Ambr 250 Perfusion: Good Match to 5 L for VCD & Titer
ViableViableCell
Cell
DensityDensity
ViableCell
Density(Mcells/mL)
cells/mL)(M(Mcells/mL)< 50%1500

ViableCell Density
Cedex
Viability(%)
Cedex
Viability(%)
Cellviability(%)(M cells/mL)

Low Capex Investment

50Bioreactor
titer
Bioreactor titre

1. Process Intensification Satisfies Key Business DriversCBioreactor titre

Prity Bengani-Lutz, Priyanka Gupta
Sartorius Stedim North America Inc., 565 Johnson Avenue, Bohemia, NY 11716

A95Ambr® n=2

92.5
Intensifying
N-1 SeedTrain is the easiest to implement with minimum change to the Fed-Batch process
5L n=1
It results
in upto 2 × higher titer, upto 50% increase in throughput & lower COGS compared to Fed-Batch
90051015203025Time calculated (days)

Intensified Stirred Tank N-1 Seed Train

CRD

2,000 L

97.5

N-2N-1 Perfusion

RM 50

STR 500 + ATF

Intensified RM 200 N-1 Seed Train (Economical)
N-2

N-1 Perfusion

Cell Culture Media (Virus Filtered)----

Seed Train Intensification
HCD cell banks | direct thaw inRocking Motion (RM)
RM Perfusion (before N-3) to skip
N-2 (Fed-Batch)
N-1 Perfusion using RM or stirredtank reactor (STR)

Main Bioreactor Intensification

Clarification Intensification

High inoculation FB from
N-1 Perfusion
Intensified Fed-Batch:a) Concentrated FB = Product inbioreactorb) (Dynamic) Perfusion = Productin permeatea) Single use centrifuge(e.g., Sartorius Ksep®)b) Multi column chromatography(e.g., Sartorius BioSMB)--

STR 2000 + ATF

Biostat® RM 50L(with the single BIOSTAT® B tower)

N-1 perfusion enables high inoculationof production bioreactor
Reduce culture length of productionbioreactor (~40%) – 17 seed train days
(25 - 29 total USP days)
Biostat RM 200L
with single tower closed lid

Biostat RM 200L
with single tower closed lid

Biostat STR Gen 3 - 500L-----RM 50
Biostat® RM 50L(with the single BIOSTAT® B tower)

3 Bioreactors & 3 steps(instead of 4 each in FB)
3 - 6 more batches / year / train

Biostat RM 200L
with single tower closed lid

RM 200 Perfusion

Upto 25% reduction in investment cost
50% lower consumables cost per batch
Upto 20% reduction in productionbioreactor culture length: 14 seed train
days (22 - 26 total USP days)

Biostat RM 200L
with single tower closed lid

Biostat® STR Gen3 - 2000L

STR 2000 + ATF

Biostat RM 200L
with single tower closed lid

Biostat® STR Gen3 - 2000L

7. Intensified N-1 Seed Train: 10 × Productivity Increase

Ref: BPOG Technology Roadmap 1st edition, 2017; 3. Process Technologies Bio© BioPhorumOperations Group, Ltd.

--Traditional Batch (Top) Versus Perfusion (Below) Seed Train Cell Culture Process Workflow at Intas

3. Clone Selection to Process Development
Save Time, Run More Experiments and Obtain Useful Data Using High Throughput, Multi-Parallel Ambr Systems®

Fast-track clone screening using Ambr® 15
Operator time spent perbioreactor for 14 day run
20

Accelerate process development using Ambr® 250

Capacity for bioreactor tests
per operator per year

Operator time spent for15 day run

1000

18.7

20Capacity for bioreactor tests
per operator per year

Hours

Hours

15600

8422

400

SS – 50 L

SS – 500 L

SS – 4,000 L

N-4

N-3

N-2

N-1

N

264

800

12Biostat® RM 10 L

300

845
16125 mL - 1,000 mL

Take Down | Clean Up

250

Sampling | Reactor
Support200

Inoculation

10Batch Readiness

150

125 mL - 1,000 mL

Biostat® RM 50 L

Bioreactor 4,000 L

N-2

N-1

N

Reduced seed train from 5 to 3 steps
saving time and cost
N-1 perfusion increased cell culture
productivity 10 × with a smaller
manufacturing footprint
Showed comparable growth kinetics
and yield with a control process at50 L scale
Target VCD reached with > 98% cell
viability in 6 – 8 days of fed-batch
cultureSource: Malla R et al., BioProcess J, 2021; 20.

132

1004

2.6

5200

1.9

50

750006 × 2 L Glass Ambr® 15
Bioreactors (24 vesselsystem)

Ambr® 15(48 vesselsystem)

6 × 2 L Glass Ambr® 15
Bioreactors (24 vesselsystem)

8. Process Scale-up & Summary

600Ambr® 15(48 vesselsystem)

Traditional Ambr® 250 Ambr® 250Glass 2 L(12 vessel (24 vessel(1 L W.V.)
system)system)

Traditional
Glass 2 L
(1 L W.V.)

Ambr® 250
(12 vesselsystem)

Ambr® 250
(24 vesselsystem)

Sartorius Upstream Process Intensification Platform: PD to Manufacturing
Clone & Media Screening | Cell Line Development

Process Development | Characterization

Clarification

4. Process Development & Characterization---- -Ambr® 250 Perfusion is Efficient & Cost Effective Compared to Traditional Benchtop Perfusion
System payback period 1 - 2 years
The major source of cost savings is media cost reduction
with Ambr® 250 High Throughput Perfusion
Annual costs were calculated for three experimentcapacity scenarios, for both:
1 × (Ambr® 250 High Throughput Perfusion – 12 vessels)
2 × (6 × 2 L benchtop bioreactors)
Annual cost savings calculated as:cost (12 × Ambr® 250) – cost (12 × 2 L)
Cumulative annual cost savings include initial CAPEX
investment

Predicted Cumulative Annual Savings withAmbr® 250High Throughput Perfusion (12-vessel)
4

Cost Savings (Millions USD)

32
10-1-2
0		

1		

2		

3

Time (Years)

45---4Cell® XtraCHO

Ambr® 154Cell® XtraCHO

Ready to Hydrate |Use Media & Buffers®

MODDE® for DOEAmbr® 250 HTUnivessel® & Biostat®

Palletanks® &Biostat® RM
BiostatSTR®
®
Flexsafe Pro Mixer (Fed-Batch & Perfusion (FB & Perfusion with ATF)Media | Buffers

Seed Train | Production

SIMCA®
for MVD

Ksep®

Sartoclear®

Sartopore® 2Sartoclear®

Sartopore® 2Clarification

Summary
Process Intensification maximizes productivity and flexibility and can be implemented step-wise or end-to-end
High throughput Ambr® systems fast-track biopharm perfusion process development saving time and money
N-1 Perfusion results in upto 2 × high titer, lower COGS and is the easiest to implement in an existing facility
PAT sensors enable culture monitoring, automated inoculation of subsequent cultures & better process control