Presented at INTERPHEX on March 21-23, 2017. Regulatory guidelines have shaped industry best practices around adventitious virus contamination risk mitigation in terms of patient safety. A recent survey indicated that the cost to remediate a virus contamination can run into millions of dollars, not including commercial impact. Today, the industry is taking a closer look at minimizing the business risk associated with viral contaminations and is taking a more holistic view of risk mitigation. This approach includes virus prevention and detection in addition to removal. From cell culture seed train to final fill vial, the discussion will describe the potential risks associated with different areas of biotech processes and what can be done to minimize adventitious virus risk in those areas. The overarching strategy of risk mitigation will include evaluation of raw materials, modified expression systems, environmental controls, upstream and downstream processing, testing and regulatory considerations.

1. Dr. Alison Armstrong
Senior Director
Global Head of Field Development Services
MilliporeSigma
Holistic Approaches To
Virus Risk Mitigation

2. Plavsic, M. BioPharm International, May 2016. p.40
The Safety TriangleThe Safety Triangle
PreventDetect
Remove
Holistic
Virus Risk
Mitigation
Raw material control
or barrier technology
Process’s ability to
mitigate potential risk
Ensure that the approach to risk works

3. Why a Holistic Approach?Why a Holistic Approach?
At the Commercial StageAt the Commercial Stage
Industry trends:
•Addressing business risk, not just regulatory
requirements
•Contaminations have happened and drive a need
for virus risk mitigation
•Not everyone has felt the impact and may
question the need
We are trying to prevent:
•Consent decree by the FDA
•Plant shutdowns and drug shortages
•Expedited approval of competitive therapies
•Public relations problems
•Costly “overhauls” of manufacturing practices

4. Why a Holistic Approach?Why a Holistic Approach?
Before the Commercial StageBefore the Commercial Stage
Industry trends:
•Virus risk mitigation implemented earlier
•Not just for commercial drugs
•Lower business risk
•Proof of concept prior to commercial
implementation
We are trying to prevent:
•Shutdowns that can last up to a year
•Delays to market
•CAPA’s that drive unplanned costs on tight
budgets
•“Knee jerk” responses to fix unplanned
problems

5. Why a Holistic Approach?Why a Holistic Approach?
Time = Planning = Savings
 Time is something you can’t get back
− Time to investigate options for short term/long term impact
− Time to engage the right business partners (CMO’s, contract testing labs, vendors)
 Time to build awareness around “safety by design”
− A unified, simple message to all areas of the company
− All personnel can be trained appropriately
 Time to avoid unwanted CAPA’s

6. Plavsic, M. BioPharm International, May 2016. p.40
Build The Safety TriangleBuild The Safety Triangle
PreventDetect
Remove

7. Where toWhere to
Assess RiskAssess Risk
People
Raw Materials
Process
Technology
Process Step
Detection
Strategy
Interlocking cogs
require integrated
approaches

8. Sources of ContaminationSources of Contamination
Exist at Multiple StagesExist at Multiple Stages
Raw
Materials
Facility registration (ISO, cGMP,
FDA), component grade, component
concentration rodent attractant
Geographical origin,
synthetic or mined,
fermented, plant or
animal derived
Added step to raw
materials manufacturing
Warehouse control, lack of
transport vehicle control

9. Low
Medium
High
Risk OutputRisk Output
RelativeRisk Glucose is a rodent
“attractant”
Each bar represents a CCM component
Raw
Materials

10. Potential Virus
Contamination
• Retrovirus, parvovirus,
herpesvirus, adenovirus
• Transmittable between
generations
• MVM
• Dormant adventitious agents
MCB Testing &
Seed Train Monitoring
• qPCR and next-gen
sequencing vs. compendial
methods
• Deep sequencing
• Dormant,
difficult to
detect
• Likely to see a
sign of infection
• MVM
Cell Line DevelopmentCell Line Development
Raw
Materials
• Interference/matrix effects
• Knowledge of target required
for increased sensitivity
• Deep sequencing – raw data
needs bioinformatics to filter
through the information

11. Mitigation ActionsMitigation Actions
Raw
Materials Eliminating or reducing the load
with a layered approach

12. Plavsic, M. BioPharm International, May 2016. p.40
Build The Safety TriangleBuild The Safety Triangle
PreventDetect
Remove
Raw materials control
begins the “prevent”
strategy

13. Process
Step
Identify the High Risk/High Impact AreasIdentify the High Risk/High Impact Areas
Downstream Goals:
Meet clearance targets,
protect product safety
Fill and Finish Goals:
Protect product safety
Upstream Goals:
Maintain “clean” MCB,
prevent ingress through
bioreactor inputs
Media and Buffer Prep Goals:
Prevent virus ingress
Assess risk based on:
 Integrates probability and impact
− Amount (High/Medium/Low)
− Impact (product safety, regulatory, business)
 Group and prioritize areas across entire process

14. Process
Technology
Current Virus Reduction TechnologiesCurrent Virus Reduction Technologies
HTST or UV:
Virus inactivation
Single Use:
Gamma irradiated
closed systems
Virus Filtration:
Size exclusion
viral clearance
Chromatography:
Multi modal viral
clearance
Low pH/Solvent/Detergent:
Viral inactivation
Closed SU Sampling:
Uncompromised sample
collection and storage
Contract Labs:
Validation, detection and
characterization

15. Process
Technology
Virus Resistant Cell lines
Prevent amplification in the
bioreactor
• Solves to the challenge of identifying known and
unknown sequences
• “A priori” sequence knowledge is not required
• Sequence of interest is readily available for rapid
development of confirmatory assays
New Technologies:New Technologies:
Where Previously There Was No SolutionWhere Previously There Was No Solution
Detection Assays
Next Generation Sequencing (NGS)
Bacteriophage T3, complete genome; NC_003298 (AJ18471)

16. Process
Technology
HTST Pre-treated Raw
Materials
Virus Filtration of
Cell Culture Media
• New solutions for heat-sensitive components
• Easy to implement
• Scalable from bench to commercial
Point-of-origin Strategy
• Some challenging components (e.g. glucose)
• Reduces risk at the source
• Supplier takes on PD challenges & validation
Point-of-use Strategy
New Technologies:New Technologies:
Where Previously There Was No SolutionWhere Previously There Was No Solution

17. Plavsic, M. BioPharm International, May 2016. p.40
Build The Safety TriangleBuild The Safety Triangle
PreventDetect
Remove
Barrier technology adds to
the “prevent” strategy
Process step and process
technology define the
“remove” strategy

18. Upstream Downstream Final Fill
Business Regulatory Product Safety
Concerns Goals
Detection
Strategy
The 3 QuestionsThe 3 Questions
Where/Why? What For? How?Where/Why? What For? How?
Where and Why:
What for:
Target agents based on
•Raw materials source and vendor documentation
•Known/past pathogens
Main “issues”
•Screens for specific current/past pathogens
•Limited detection to a small number of known pathogens
•Ignores emerging, novel viruses

19. Detection
Strategy
How:
Considerations for
Method Selection
• Sensitivity of assay limits detection
‒ All assays have a LOD; sample volume limitations
• Cell lines may not be permissive for some known or novel viruses
• Interference/matrix effects
• Anti-virus antibodies in FCS used in in vitro assays
• Cytotoxicity of indicator cells
• Inhibition of PCR assay enzymes
The 3 QuestionsThe 3 Questions
Where/Why? What For? How?Where/Why? What For? How?

20. Plavsic, M. BioPharm International, May 2016. p.40
Build The Safety TriangleBuild The Safety Triangle
PreventDetect
Remove
Where and what to
sample for starts the
“detection” strategy

21. People
The Touchpoint of Every CogThe Touchpoint of Every Cog
Safety by Design
Develop a Virus
Safety Culture
• Proper operator training
• Proper thinking, “do the right thing”
• Stay home when you are sick!
• Well designed processes minimize human error
• Certain process technologies can reduce or eliminate the people
factor (such as single use technology with sterile connectors or
pre-weighed, closed powder delivery)

22. PreventDetect
Remove
People complete
the strategy
Plavsic, M. BioPharm International, May 2016. p.40
The Safety Triangle BuiltThe Safety Triangle Built

23. Interpreting the AssessmentInterpreting the Assessment
CategoriesCategories
Risk Assessment
Product
Safety
Regulatory
Strategy
Business
Risk

24. • Not negotiable
• Risk is absolutely minimized
• Detection strategy confirms product safety
Interpreting the AssessmentInterpreting the Assessment
Product Safety
Regulatory Strategy
• STRATEGIZE: Orthogonal and layered
• PROCESS: Designed in a way to minimize non-conformance
• TECHNOLOGY: Robust validated LRV through process upsets

25. Interpreting the AssessmentInterpreting the Assessment
Business
Risk
Quantitative Approach
Contamination cost hard to model
•Multiple variables
‒ plant down time
‒ cost of a CAPA
•Hard to quantify “soft” impacts
‒ damage to reputation
‒ delay in filing
‒ hosting regulatory visits
Final Strategy Integrates Both Approaches

26. Raw Materials Testing Lot Release and Biologic Support Testing
Controlled Raw Materials
Collaborate With Your Vendor to Reach Your GoalsCollaborate With Your Vendor to Reach Your Goals
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

27. Example: Protect the BioreactorExample: Protect the Bioreactor
CCM and
Supplements
Operators
Cells
Bioreactor
Hardware
Low
High
Medium
Buffers, Sparge
and Overlay Gas
Sampling
Risk/Impact Rating
Detection
Strategy

28. The Safety TriangleThe Safety Triangle
PreventDetect
Remove
Holistic
Virus Risk
Mitigation
Plavsic, M. BioPharm International, May 2016. p.40