Watch the on demand recording here: https://bit.ly/3cHnFzP All biologics production processes focus on preventing adventitious microorganisms from entering upstream processes. Regulatory guidance for upstream processes focuses on comprehensive testing and characterization of raw materials. While there is no specific mandate for implementing viral clearance technologies, manufacturers are increasingly implementing practices that mitigate the risk of contamination and the consequent disruption to manufacturing operations. This webinar will discuss the potential sources of viral contaminants, the strengths and limitations of different detection and removal technologies and explain how they can be integrated into a comprehensive, holistic, upstream viral safety strategy. In this webinar, you will learn about: - How integrating multiple approaches can reduce contamination risks - Retention performance with different filters for cell culture media processing

1. The life science business of Merck KGaA,
Darmstadt, Germany operates as
MilliporeSigma in the U.S. and Canada.
A Holistic
approach to
Upstream Viral
Safety
Craig Jackson, Senior Product Manager
5 March 2020, Bedford, MA

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

3. Agenda
1
2
3
Background
Process Step
Raw Materials
4
5
Mitigation Technology
Detection Strategy

4. Viral Risk Management Requires an Integrated Approach
Holistic Upstream Viral Safety | 5 March 2020
People
4

5. Holistic Upstream Viral Safety | 5 March 2020
Understanding Your Viral Contamination Risk Profile
Risk Perception
How does your company
evaluate risk?
Risk Tolerance
Does information change your
behavior?
5

6. Reported Major Viral Contamination Events
Holistic Upstream Viral Safety | 5 March 2020
Viral Contamination
6

7. Elements of a holistic approach
Holistic Upstream Viral Safety | 5 March 2020
Process’s ability to
mitigate potential risk
Ensure Safety of Raw
Materials and Processes
Ensure that the approach to risk works
7

8. Low probability but High Impact
Viral Contamination
Holistic Upstream Viral Safety | 5 March 2020
Precommercial
• Facility shutdown can last up
to a year
• Delays to market
• CAPAs – unplanned costs
• ‘Knee jerk’ responses to fix
unplanned problems
Commercial
• Consent decree by FDA or EMA
• Plant shutdowns & potential
drug shortages
• Expedited approval of
competitive therapies
• Public relations problems
• Costly overhauls of
manufacturing practices
8

9. Improved Planning Avoids Costly Contamination Events
Holistic Upstream Viral Safety | 5 March 2020
• To investigate
• Find right business
partners to help
• Build company
awareness
• Avoid unwanted CAPAs
Save time….
9

10. Improved Planning Avoids Costly Contamination Events
Holistic Upstream Viral Safety | 5 March 2020
• To investigate
• Find right business
partners to help
• Build company
awareness
• Avoid unwanted CAPAs
Trends
• Risk mitigation implemented earlier
• Focus on reducing business risk, not just
meeting regulatory requirements
• Increased awareness of the need for virus
risk mitigation plans
• Increasing consideration of newer
risk reduction technologies
Save time….
10

11. Risk Management Requires an Integrated Approach
Holistic Upstream Viral Safety | 5 March 2020
People
11

12. Different Considerations
Holistic Upstream Viral Safety | 5 March 2020
Upstream
Biosafety reduces business risk
Guidance documents focus on raw
material testing
Downstream
Regulatory requirements for bacteria
and virus removal
Virus Safety Upstream vs Downstream
Process
Step
12

13. Multiple Potential Sources of Viral Contamination
Holistic Upstream Viral Safety | 5 March 2020
CCM and
Supplements
Operators
Cells
Bioreactor
Hardware
Facility &
Utilities
Sampling
Detection
Strategy
Upstream
Process
Step
13

14. Risk Management Requires an Integrated Approach
Holistic Upstream Viral Safety | 5 March 2020
People
14

15. Holistic Upstream Viral Safety | 5 March 2020
Reduce or Mitigate Risk Where Possible
Raw Materials
Replace animal-derived
components in media
• Lower risk recombinant
alternatives (r-Insulin,
r-Trypsin, etc)
• Source from lower risk
geographies – 9CFR testing
Adopt chemically-
defined, animal-
derived component-
free media
• Informed product
selection: quality,
documentation, services
Implement pretreated
high-risk materials
• Gamma irradiated
• Supplier-treated
materials offer additional
security
Raw
Materials
Informed Product Selection:
• Select the right product for intended use
• Select business partners with transparent quality programs
15

16. Holistic Upstream Viral Safety | 5 March 2020
Risk Profile is the Sum of Risk
Viral Risk Assessment
Raw
Materials
Material and
Source
Manufacturing
Facility
Repacking of
Material
Concentration in
Media
Risk Profile
USP
EP
JP
16

17. Holistic Upstream Viral Safety | 5 March 2020
Risk Profile is the Sum of Risk
Viral Risk Assessment
 Geographical origin
 Source: synthetic or
mined, fermented, plant
or animal derived
• Facility registration (ISO,
cGMP, FDA)
• Component grade,
concentration
• Supply chain
• Rodent control/attractant
• Added step to
manufacturing
• Increases exposure
• Control during transport
and in warehouse
• Not limited to one
component
• Consider in context of all
raw materials
Material and
Source
Manufacturing
Facility
Repacking of
Material
Concentration in
Media
Risk Profile
USP
EP
JP
Raw
Materials
17

18. Holistic Upstream Viral Safety | 5 March 2020
Risk Profile Cell Culture Media
Viral Risk Assessment
USP
EP
JP
USP
EP
JP
Low score Higher score
Material and
source
 Countries with robust and
well established quality
Systems
 Synthetic/mined
 Pharma grade
 Low risk rodent exposure
 Countries with emerging
Quality Systems
 Animal derived
 Industrial or <98% purity
 High risk rodent exposure
Repacking
 No repacking
 cGMP facility
 Low risk rodent exposure
 Material repacked
 No/unknown registration
 High risk rodent exposure
Facility
 cGMP  No/unknown registration
Concentration
<1 mg/L >1000 mg/L
score
score
score
score
+
+
+
=
Final
Risk
Score
Raw
Materials
18

19. Holistic Upstream Viral Safety | 5 March 2020
Risk Profile Cell Culture Media
Viral Risk Assessment
USP
EP
JP
Assess risk of each component
 Concentration
 Repackaging
 Facility registration
 Origin score
Plot relative risk
• Identify high risk components
Develop mitigation plans
• Address driving forces behind
scores
Raw
Materials
19
IndividualCCMcomponentsl

20. Holistic Upstream Viral Safety | 5 March 2020
Point-of-Origin Treatment for Glucose
High Temperature Short Time (HTST)
• HTST highly effective viral inactivation technology
• ≥6 log MVM reduction ~ @ 102°C with a ≥10 sec RT
 Suitable for challenging-to-filter solutions
 Supplier takes on HTST challenges:
caramelization, precipitation, burn-on
 Reduces risk at source: reduces the
likelihood of contaminated materials
entering the plant
Condition (Temperature/time (secs))
Raw
Materials
20

21. Holistic Upstream Viral Safety | 5 March 2020
Risk Management Requires an Integrated Approach
People
21

22. Single-Use Technologies Enable Closed Processing
Holistic Upstream Viral Safety | 5 March 2020
Mitigating Viral Contamination Risk
SamplingBioreactorsMixers
• Single-use, gamma-sterilized components enable closed processing
• Reduce contamination risk, increase speed and manufacturing flexibility
• Minimizes impact in event of contamination
Connectors
Mitigation
Technology
22

23. Holistic Upstream Viral Safety | 5 March 2020
Why?
Point-of Use Treatments
USP
EP
JP
 Cell culture media are complex
with many components
 Point-of-origin treatment may not
be feasible for every component
USP
EP
JP
RelativeRisk
Concentration Repacking Facility Reg Origin & Source
Mitigation
Technology
23
Individual CCM componentsl

24. Holistic Upstream Viral Safety | 5 March 2020
On what?
Point-of-Use Treatments
USP
EP
JP
Point-of-use treatments are the last
line of defense against contamination
USP
EP
JP
RelativeRisk
Concentration Repacking Facility Reg Origin & Source
Cumulative risk of
‘low and medium risk’
can be high
Mitigation
Technology
24
Individual CCM componentsl

25. Holistic Upstream Viral Safety | 5 March 2020
HTST Treatment of CCM
Point-of-Use Treatments
Strengths:
• Broadly effective
• Time proven
Weaknesses:
• Large capital investment needed
• No simple small-scale systems
• Large footprint, needs CIP
• Some CCM components are heat labile
 Visible: Precipitation, caramelization
 Invisible: Impact on cell culture performance
Mitigation
Technology
25

26. Holistic Upstream Viral Safety | 5 March 2020
Evolving Filtration Technology for CCM
Point-of-Use Treatments
Micro-organism
Challenges for traditional
sterilizing–grade filters
Spirochetes,
mycoplasma,
Spiroplasma
Some passage through 0.1
and 0.2 µm filters
Viruses Not removed
0.45 µm
0.2 µm
0.1 µm
Virus
retentive
Mitigation
Technology
26

27. Potentially
fouling
components
Large components
 Recombinant insulin
 Dextrans
Poorly soluble components
 Salts of trace metals
Poloxamer
 Poloxamer 188 is included in
many cell culture media as a
cell shear protectant
 Known to cause virus filter
fouling
Hydrophobic components
 Fatty acids
 Steroids (hydrocortisone,
cholesterol)
Holistic Upstream Viral Safety | 5 March 2020
Point-of-Use Treatments
Hard to filter components in CCM
Mitigation
Technology
27

28. Holistic Upstream Viral Safety | 5 March 2020
Virus Filters for CCM Outperform Downstream Virus Filters
Point-of-Use Treatments
Mitigation
Technology
28

29. Holistic Upstream Viral Safety | 5 March 2020
Microbial Retention on Different Filters
Point-of-Use Treatments
High Bubble Point
0.1 µm PVDF
0.1 µm PVDF 0.22 µm PVDFViresolve® Barrier
Filter
LogReductionValue(LRV)
Complete retention
Microorganism
L. illini
Width (µm) Length (µm)
0.1 171
Microorganism
1 Bergey’s Manual of Systematic Bacteriology
Complete retention
Upstream Virus Filters Protect Against Spirochete (Leptonema illini) contamination
Mitigation
Technology
29

30. Holistic Upstream Viral Safety | 5 March 2020
Upstream Virus Filters Retain Bacteria & Virus
Point-of-Use Treatments
Largest Smallest
Virus filters designed for cell culture media provide protection against
virus and microbial contaminants not removed by sterilizing filters
Mitigation
Technology
Complete retention
30

31. Holistic Upstream Viral Safety | 5 March 2020
Cell Culture Performance Maintained
Point-of-Use Treatments
Cell culture performance
• Viable cell density
• Protein titer
• Aggregate analysis
• Glycan composition
• Charge heterogeneity
Media analytics
 Mass spec
 Amino Acid HPLC
 Soluble vitamin HPLC
 NMR
 ICP-OES
Viable Cell Density
Glycosylation Patterns
Mitigation
Technology
31

32. Holistic Upstream Viral Safety | 5 March 2020
Point-of-Use Technologies Reduce Contamination Risks
Point-of-Use Treatments
1
2
3
Virus filters designed for CCM
HTST treatment of CCM
Single-use components
0.1 or 0.2 µm
filtration of cell culture
media and feeds
Then Now
Mitigation
Technology
32

33. Holistic Upstream Viral Safety | 5 March 2020
Risk Management Requires an Integrated Approach
People
33

34. Holistic Upstream Viral Safety | 5 March 2020
Where, Why, and What for?
Traditional Viral Detection
General Process Design
Final Fill:
Product safety
Downstream:
Regulatory
requirements
Upstream:
Reduce
contamination
risk
Strengths:
• Time-proven
• Solid baseline for ‘clean’ raw
materials
Limitations:
• Long time to results
• Limited test method sensitivity & specificity:
“You only find what you are looking for”
• Not compatible with continuous processing
Detection
Strategy
34

35. Holistic Upstream Viral Safety | 5 March 2020
No longer a rate–limiting step
Next-Generation Viral Detection Methods
New Platforms:
Broad based virus
screening with the
sensitivity of PCR
• Virus testing is no longer a rate-limiting step: ‘days not weeks’
• Offers sensitivity and specificity
• Reduces animal testing
Detection
Strategy
Automated nucleic
acid extraction of
CHO Bulk harvest
material
Positive controls
spiked into sample
Highly multiplexed
degenerate PCR
amplification
Broad coverage of
over 5000 virus
variants
Detection sizing of
amplified products
Equivalent
sensitivity to
existing PCR
methods
35

36. Holistic Upstream Viral Safety | 5 March 2020
Risk Management Requires an Integrated Approach
People
36

37. Holistic Upstream Viral Safety | 5 March 2020
Work with your Suppliers
Summary: Integrated Viral Safety
Raw Materials Testing
Lot Release and Biologic Support
Testing
Controlled Raw Materials
Expression
System and
Cell Line
Development
Upstream Viral Mitigation
Technology
Contract Biologic Manufacturing
Purification Technology With Viral
Clearance
Downstream Viral Clearance
Validation
Downstream Process Development
and OptimizationUpstream
Process
Development
37

38. Craig.Jackson@emdgroup.com
Craig Jackson
Thank you!
Questions?
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