Watch the interactive recording here: https://bit.ly/2OdLYwX Process optimization and upstream intensification led to smaller, more efficient biomanufacturing facilities becoming more commonplace, with smaller facilities comprised primarily of single use or hybrid technology capable of producing significant amounts of drug product. Such changes, however, bring new challenges, like managing the supply of huge amounts of cell culture media or buffers within smaller footprints. In this webinar two topics will be addressed that help to intensify upstream and downstream processes and address the challenges of future facilities. Bulk powders of cell culture media (CCM) or single chemicals often show physical disadvantages. CCM powders with fine particles show high dust formation and poor flowability. In addition, dissolution is time consuming due to floating of light particles on the water surface. For the pursued intensification of upstream processing, media preparation times are becoming a serious bottleneck. This mainly accounts for the much higher media consumption or higher concentrated media formulations for future continuous upstream processes. Granulated material can overcome limitations with CCM powder, while additionally being a viable option to reduce caking of bulk chemicals like buffers. Buffer production for downstream processing remains a significant portion of the facility footprint, labor needs and equipment cost. As downstream operations are essentially product-mass-based, increased productivity in upstream will lead to a proportional increase in demand for downstream buffers. Merck KGaA Darmstadt, Germanty R&D has their expertise in concentration of buffers to improve and streamline buffer management.

1. The life science business of Merck KGaA,
Darmstadt, Germany operates as
MilliporeSigma in the U.S. and Canada.
Process Intensification for
Future Bioprocessing
Corinna Merkel
Scott Wilson
July 25th 2019

2. The life science business
of Merck KGaA, Darmstadt,
Germany operates as
MilliporeSigma in the U.S.
and Canada

3. 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
325.07.2019 Process Intensification for Future Bioprocessing

4. Next Generation Processing:
Evolutionary Journey in mAb Manufacturing
End state
Today
Near-term
Mid-term
Process
Batch
Intensified
Connected
Continuous
Format
Stainless
Hybrid
Single Use
Single Use
Closed
Systems
Our Mission
Developing technologies,
applications and expertise
that enable biotherapeutic
manufacturers to confidently
enter the era of next
generation processing.
Less footprint
Less capital
Reduction in
construction time
Reduction in OPEX
90%
90%
70%
60%
In Numbers
425.07.2019 Process Intensification for Future Bioprocessing

5. Next Generation Processing:
Evolutionary Journey Across Many Disciplines
1
Intensified
Upstream Technologies
Buffer Management
Downstream
Continuous Capture
32
FACTORY
OF THE
FUTURE
525.07.2019 Process Intensification for Future Bioprocessing

6. Upstream Process
intensification through
roller compaction
Corinna Merkel

7. Process Intensification for Future Bioprocessing7
Preparation time for hydration of
cell culture media powder
• High media demand for perfusion processes
• Highly concentrated cell culture media
Pain points for cell culture
media powder
Foot print for storage of dry
powder media
Can be addressed by granulated cell culture media

8. What are granulated cell culture media or granulated single
chemicals?
Process Intensification for Future Bioprocessing8
1 mm
Difference between bulk and granules is physical format only
Cell culture media
powder
Cell culture media
granules
Single chemicals
bulk powder
Single chemicals
granules

9. Introduction to roller compaction
Process Intensification for Future Bioprocessing9
•Granulation technology
•No water or excipient is added to
the process
•Works with compression force
only
•Main process parameters:
−Compression force
−Rotor mill grid size
−Powder characteristics
0 0 0 0
0 0 0
0 0
0
0
0
0 0
0
0
0
0
Powder
Plate
Granules
Powder is filled in
funnel
Powder is
conveyed to rolls
by mixer
Powder is
compressed by
rolls
0 0
0
0
0
0
000
Plate is formed
Plate is milled to
granules
Vibrating sieve
separates granules
from fine particles
Fine particles
recirculated
GMP roller compactor RC120 (Powtech)
Recirculation
of fines
Rolls
Vibrating sieve
Rotor mill
9

10. Compacted Chemicals
103669 Glycine Granules EMPROVE® EXPERT
More launches in 2019/2020
Advantages
• Less caking
• Improved handling
Compacted Cell Culture Media
• 103687 Cellvento® CHO-220 COMP
• 103688 Cellvento® Feed-220 COMP
• 103795 Cellvento® 4CHO COMP
• 103796 Cellvento® 4Feed COMP
Advantages
• Faster dissolution
• Less storage capacity, better flowability,
less dust
• Same cell culture performance
Current products using compaction technology
Process Intensification for Future Bioprocessing10
On-demand Webinar:
Efficient Handling of Chemical Raw Materials in
Biopharmaceutical Manufacturing by Dr. Raphael Gübeli

11. Dissolution kinetic of Cellvento® 4Feed (25°C)
11 Process Intensification for Future Bioprocessing
7 min 33 min 60 min
Powder
3 min 5 min
Granules
Process intensification by decreasing hydration time of cell culture media.
Critical for high media demand in perfusion or next generation
manufacturing of concentrated media.
Advantages of compacted media/feed
Decreased dissolution time
T i m e [ m i n ]
Normalizedconductivity
0 1 0 2 0 3 0 4 0 5 0 6 0
0 . 0
0 . 5
1 . 0
G r a n u l e s
P o w d e r
9 - f o l d
t i m e p o i n t o f v i s u a l
d i s s o l u t i o n

12. Advantages of compacted media/feed
Lower bulk volume → less storage/transport space needed
Process Intensification for Future Bioprocessing12
Save up to 1/3 of storage space with
granulated cell culture media
Smaller pack sizes may reduce footprint
in future manufacturing facilities.
P
o
w
d
e
rG
r
a
n
u
le
s
P
o
w
d
e
rG
r
a
n
u
le
s
0
1
2
3
B u l k v o l u m eBulkvolume[L/kg]
C H O - 2 1 0 F e e d - 2 1 0
- 3 1 %
- 2 8 %

13. Advantages of compacted media/feed
Decreased dust formation
Process Intensification for Future Bioprocessing13
Online Aerosol Spectrometer (Grimm)
Measures dust fractions from 0.265-34 µm
Time and cost savings by:
• Improved handling
• Reduced risk of contamination
• Less cleaning efforts
• Increased safety for employees
C e l l v e n t o  C H O - 2 1 0
P
o
w
d
e
r
G
r
a
n
u
le
s
0
1 1 0
0 7
2 1 0
0 7
3 1 0
0 7
4 1 0
0 7
P o w d e r
G r a n u l e s
counts/L * * *

14. Improved flowability – better handling
Process Intensification for Future Bioprocessing14
Time savings with convenient handling of
granulated material: weighing and
dispensing into hydration vessel.
Powder Analyzer
Avalanche
angle
Advantages of compacted media/feed
1
3 0
4 0
5 0
6 0
7 0
8 0
F l o w a b i l i t y
AvalancheAngle[°]
P o w d e r
G r a n u l e s
N o n - s a t i s f a c t o r y
A v e r a g e
G o o d
E x c e l l e n t

15. 15
Compaction does not impact homogeneity of media
amino acids and vitamin analytics
Process Intensification for Future Bioprocessing
• Raw materials are homogeneously distributed
• Compaction does not impact critical raw materials such as vitamins
A m i n o a c i d s
H
is
S
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r
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g
T
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G
lu
A
s
n
6 0
8 0
1 0 0
1 2 0
1 4 0
%oftheory
G r a n u l e s
P o w d e r
V i t a m i n s
F
o
lic
A
c
id
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ib
o
fla
v
in
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io
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ita
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6 0
8 0
1 0 0
1 2 0
1 4 0
%oftheory
P o w d e r
G r a n u l e s

16. Process Intensification for Future Bioprocessing16
Roller compaction does not impact pH, osmolality and
solubility
Granulated medium has same pH, osmolarity and solubility like powder
C e l l v e n t o  C H O - 2 1 0
G
r
a
n
u
l e
s
P
o
w
d
e
r
0
2
4
6
8
pH
C e l l v e n t o  C H O - 2 1 0
G
r
a
n
u
l e
s
P
o
w
d
e
r
0
1 0 0
2 0 0
3 0 0
4 0 0
Osmolality[mOsm/kg]
C e l l v e n t o  C H O - 2 1 0
G
r
a
n
u
le
s
P
o
w
d
e
r
0
1
2
3
Turbidity[NTU]
16

17. Compaction does not impact CHO cell performance
and product quality*
Process Intensification for Future Bioprocessing17
Producttiter[mg/l]
VCD[106viablecells/ml]
Powder medium + powder feed
Compacted medium + compacted feed
Same VCD, product titer and glycosylation profile
between powder and compacted medium and feed
*Customer data
Glycans[%]
Glycosylation
Viable cell density
Product titer

18. Advantages of granulated single chemicals
Why is compaction of single chemicals beneficial?
Process Intensification for Future Bioprocessing18
25 °C/60%
40 °C/75%
GranulesPowder
Main advantage: less caking
Cost and time savings with improved handling and increased employee
safety due to less elaborate de-caking of caked bulk material
18

19. Summary
Process intensification by granulated materials
Process Intensification for Future Bioprocessing19
Lower Bulk
Volume
Elevated
Dissolution
Speed
Improved
Flowability
Reduced Dust
Formation
Reduced
process time
Reduced footprint
Improved
handling
Reduced contamination
risk & improved
employee safety
BulkGranulated
material
Less Caking
Improved
handling &
employee safety
Less
Segregation
Improved
homogeneity
Cell culture media
Single chemicals
19

20. De-bottlenecking the
Downstream Process:
Buffer Concentrates
Scott Wilson

21. Buffer preparation is an essential function
Process Intensification for Future Bioprocessing21
1
Buffer Prep Is a Core Operation
Drives plant schedule and capacity
2
Critical, but not Value-Added
Imbalance between resources &
footprint required compared to
simplicity of buffer prep
Requirements:
5-10 L buffer per L bioreactor
15-18 unique downstream buffers
Critical attributes for process performance
Large amount of classified floor space
Significant labor requirements
How do we provide the right buffer at the right time and
specifications while minimizing our labor and footprint?

22. Facility
type
Traditional Hybrid Next Generation
BioR
volume
10,000 L 2,000 L 200 L (x3)
Titre 1g/L 5g/L 10g/L
Runs/year 15 15 30
Drug Qty 150 Kg 150 Kg 180 Kg
Decreasing Manufacturing Footprint
Maintaining Production of Drug Product
Impact of Facility Throughput versus Buffer Demand
0
50
100
150
200
250
300
350
400
450
500
0 50 100 150 200 250 300
BufferVolumeKL
Drug Product kgs
Annual Buffer Volume, L
Buffer demand proportional to drug product
produced
@150 kg MAb
> 200 kL / year
>16 kL / month
Process Intensification for Future Bioprocessing22

23. Made in
house
(MIH)
Buffer
concentrates
(CONC)
Ready-to-use
liquid buffers
(RTU)
Sourcing Preparation Protection SamplingTransfer and
Storage
Buffer Management Strategies
Process Intensification for Future Bioprocessing23

24. Impact of Buffer Preparation Method on Facility Design
Process Intensification for Future Bioprocessing24
Reduction in cleanroom area utilizing buffer concentrates
On-Site Buffer Preparation
970 m2 manufacturing cleanroom

25. Impact of Buffer Preparation Method on Facility Design
Process Intensification for Future Bioprocessing25
18% reduction in cleanroom area utilizing buffer concentrates
Buffer Concentrates
791 m2 manufacturing cleanroom
18% cleanroom footprint
Buffer concentrates shrink new facility footprint or can increase
capacity of existing facilities to free up more working space

26. Buffer Concentrates 2019
2,000 L Bioreactor Facility, 12 Runs per Year
Buffer Concentrates Compared to Made-in-House Buffers
Buffer Concentrates significantly reduce labor, consumables and QC Cost
27,23%
18,30%
31,36%
0,99%
13,15%
3,90%
2,05% 3,01%
% Total Cost MIH 2000L 5g/L
3,12%
82,77%
8,11%
0,25%
0,73% 2,74%
1,54%
0,74%
% Total Cost Conc 2000L 5g/L
26

27. Technical Assessment
Typical stock buffer concentrates
Process Intensification for Future Bioprocessing27
• >25 buffers evaluated to determine concentration factor
• 19 most commonly used buffer in Mab Processes
• Specific buffers recommended for Eshmuno® and Fractogel® resins
• Tested maximum solubility (typically 10-30x)
• Conducting stability studies in Mobius® assemblies
• ISTA ship tested (for robust shipment of sterile liquids)
Conclusions
Buffer concentrates help drive:
• Reduction in labor costs
• Reduction in facility CAPEX and overhead
• Improved operator safety
• Reduction in operational footprint
• Improved sustainability
Cost models can be customized to individual
process and facility
Some Typical Buffers Evaluated
0.1M acetate + 0.5 M NaCl, pH 5.5
1.0 M tris, pH 8.4, pH 8.5
0.05M Tris, 0.5M arginine, pH 8.4, pH 8.5
50-100 mM acetic acid, pH 3.2
50-100 mM citric acid, pH 3.2
50mM phosphate pH 7.4
0.1M citrate, pH 5.4, pH 5.5

28. 28
Control Strategies
pH control
 Sensor Accuracy, ≥ 1%
 In addition:
o Tendency to drift
o Memory effect often observed
o Frequent calibration required
Conductivity control
 Sensor Accuracy, ≥ 1%
 In addition:
o Frequent calibration required
Highest accuracy in buffer dilution with MilliporeSigma/YMC buffer dilution systems
Volumetric flow control delivers accuracy and precision for consistent
buffer preparation

29. 29
Control Strategies
pH control
 Sensor Accuracy, ≥ 1%
 In addition:
o Tendency to drift
o Memory effect often observed
o Frequent calibration required
Conductivity control
 Sensor Accuracy, ≥ 1%
 In addition:
o Frequent calibration required
Volumetric Flow Control
Flow meters and pumps are more accurate and
stable than pH or conductivity detectors
17 and 33 LPM systems
Lewa Ecodos® pump with Intellidrive®
− Volume Flow Control Accuracy <1.0%
− Highest accuracy in dilution at 20:1 ratio
or greater
Highest accuracy in buffer dilution with MilliporeSigma/YMC buffer dilution systems
Volumetric flow control delivers accuracy and precision for consistent
buffer preparation
Accurate buffer concentrate preparation and dilution negates need for pH
or conductivity conditioning

30. Facility Floor View
Process Intensification for Future Bioprocessing30
BioContinuum™ Buffer Delivery Platform
Process-Specific
Buffer Concentrates Buffer Dilution
System
Single-Use
Assemblies and
Containers

31. Summary
31 Buffer Concentrates 2019
18% decrease in footprint
dedicated to buffer preparation
With prepared concentrates labor
focus can shift away from buffer
preparation and testing.
Delivery platform can lead to
accurate buffer delivery and
increased flexibility
Reduce labor,
consumable, and QC
Costs
Develop an ‘user
defined’ inline dilution
system
Reduce Warehouse
Footprints
‘Made-in-House’ vs. Buffer concentrates