The document discusses the evolution of biosafety testing methods for biopharmaceutical manufacturing. It outlines how current batch-based manufacturing relies on biosafety testing across the entire process, including adventitious agent testing that can take weeks. It identifies drivers for change, such as novel therapies like CAR-T that require faster testing timelines, and emerging technologies like next generation sequencing, mass spectroscopy, and Raman spectroscopy that could enable more rapid and online monitoring. The document argues that internal quality systems are currently the main barrier to adoption of new testing methods, but that regulatory acceptance of alternatives and ethical obligations to reduce risk will continue pushing the industry toward more rapid molecular techniques and enclosed manufacturing approaches.

1. The life science business of Merck KGaA, Darmstadt, Germany
operates as MilliporeSigma in the U.S. and Canada.
Evolution of
Biosafety testing
to meet the needs
of manufacturers
tomorrow
Adam Inche, DPhil
Biosafety Testing Services
March 12, 2020

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

3. Agenda
Biosafety testing of today
The drivers for change
Biosafety testing of tomorrow
1
2
3

4. Biosafety Testing Evolution | Adam Inche | 12March20204
Isoenzyme, 2015 – a lesson on change from the recent past
• Enzyme isoforms to identify
cell species
• Example test in ICH Q5A for
Cell Line Characterization
• Innovative Chemistry sole
supplier to industry
Dec ‘14 Innovative
Chemistry become
unresponsive
Feb ‘15 Customers
made aware of
supply issues
Mar ‘15 Customers notified
of Isoenzyme withdrawal –
alternative ID assay offered
Aug ‘15 Launch of
CO1 Barcode assay
Mar ’16 Pass x00
samples run on CO1
Barcode platform
201620152014
• Change is never easy
• Rapid change can be
very painful

5. What do you see as the main barrier for adopting new methods in biopharmaceutical manufacturing?
• No regulatory drive
• Lack of appropriate technology
• Internal change process too difficult
• No pressing need to change
Biosafety Testing Evolution | Adam Inche | 12March2020
Poll Question 1
5

6. Biosafety Testing Evolution | Adam Inche | 12March20206
Current manufacturing processes utilize batch operations
that require biosafety testing across the whole process
Chromatography
Protein A
Bioreactor
Chromatography
Purification
Cell Line
Media and
Buffer Prep
Viral
Inactivation
Viral
Filtration
Final Fill
Cell
Expansion
Formulation
Bulk Harvest
Lot Release Testing
Drug Substance/
Final Product
Lot Release Testing
Cell Line
Characterization
Specific focus on
adventitious agents

7. Biosafety Testing Evolution | Adam Inche | 12March20207
Traditional methods for adventitious agent testing rely on
biological amplification which can take several weeks
Cell Line Characterization Bulk Harvest Lot Release

8. Biosafety Testing Evolution | Adam Inche | 12March20208
If rapid bulk harvest release methods exist today why
don’t people use them?
Incumbent BHLRT Package Rapid Alternatives
Lack of rapid
alternative to
in vitro screen
Documentation?
Anchor
to
current
methods

9. Biosafety Testing Evolution | Adam Inche | 12March20209
The drivers of change in biosafety testing of biologics are
diverse but not so different from other industries
CHANGE
Regulatory
Resource
availability
Ethical
Manufacturing
Modes
Treatment
Modalities
Technological

10. Biosafety Testing Evolution | Adam Inche | 12March202010
Regulatory documents on biosafety testing
have always been open to alternative methods
“…regarded as assay protocols
recommended for the present,
but list is not all-inclusive or
definitive.”
“…proposal for alternative
techniques…may be acceptable.”
“…PCR may be appropriate for
detection of...human…as well as
for other specific viruses”

11. Biosafety Testing Evolution | Adam Inche | 12March202011
Regulatory documents on biosafety testing
have always been open to alternative methods
“…regarded as assay protocols
recommended For the present,
but list is not all-inclusive or
definitive.”
“…proposal for alternative
techniques…may be acceptable.”
“…PCR may be appropriate for
detection of...human…as well as
for other specific viruses”
Update to
ICH Q5A
(R2) in
2020

12. Biosafety Testing Evolution | Adam Inche | 12March202012
The industry is moving to alternative
methods to fulfil its ethical obligations
Typical Cell Line Characterization Package
BioReliance® Blazar™ Rodent Virus Panel, a PCR
based alternative to antibody production test
NGS replacement,
well characterized
CHO cells

13. Biosafety Testing Evolution | Adam Inche | 12March202013
Novel therapies provide novel challenges
to existing biosafety testing strategies
Autologous
CAR-T Therapy
Characterization
& Testing
Characterization & Testing
• 4-6 weeks for full biosafety
testing
• NGS, PCR and rapid sterility
• Risk assessment for patient
infusion
Current drivers for testing:
speed, volume,
compatibility

14. Biosafety Testing Evolution | Adam Inche | 12March202014
Novel therapies provide novel challenges
to existing biosafety testing strategies
Autologous
CAR-T Therapy
Characterization
& Testing
Characterization & Testing
• 4-6 weeks for full biosafety
testing
• NGS, PCR and rapid sterility
• Risk assessment for patient
infusion
Current drivers for testing:
speed, volume,
compatibility

15. Biosafety Testing Evolution | Adam Inche | 12March202015
Next Generation Sequencing is now a firmly
entrenched technology in biosafety testing
• Unbiased, unselected analysis
• Difficult and complex samples
• Direct identification
• Relatively rapid
• Significant industry interest
(AVDTIG)
• Known to regulators
• Sensitivity can be variable
• Higher complexity sample
prep and analysis
• Detecting unknowns can still
be problematic – e.g.
rhabdovirus in SF9
• Perhaps not fast enough for all
applications
Advantages Challenges

16. Biosafety Testing Evolution | Adam Inche | 12March202016
PCR still has a lot to offer...
qPCR digital PCR
• Well accepted for specific detection
• Highly sensitive – 10GC
• Rapid
• Doesn’t discriminate live/dead
• Only detects what you are looking
for
degenerate PCR
• Targeted to virus
families
• Identify related
unknown viruses,
e.g. MKPV

17. Biosafety Testing Evolution | Adam Inche | 12March202017
Novel therapies provide novel challenges
to existing biosafety testing strategies
Autologous
CAR-T Therapy
Enclosed Process
• Reduce/eliminate
risk of introduction
of adventitious
agents
• Risk-based testing
strategy - blood
transfusion approach
Testing still
required

18. Biosafety Testing Evolution | Adam Inche | 12March202018
Moving towards process intensification will
change biosafety testing needs
Batch Process
Intensified semi-continuous process
Continuous process
Increased timeline
pressure:
• Shipping takes days
• Cultivation assays
take several weeks
Driving towards:
• Faster molecular methods
• Testing closer to mfg line

19. Biosafety Testing Evolution | Adam Inche | 12March202019
Monitoring of the bioreactor will be critical
in intensified processes
On-line
In-line
At-line Near-line

20. Biosafety Testing Evolution | Adam Inche | 12March202020
Monitoring of the bioreactor will be critical
in intensified processes
NGS
Mass Spec
Data challenge remains
• How to connect output
to biosafety risk?
• How to detect latent
risks?
Raman
Spectroscopy

21. Biosafety Testing Evolution | Adam Inche | 12March202021
How are we driving change in biosafety testing?
Available Now
BioReliance® Blazar™
Rodent Panel Supplementing /
replacing in vitro assay
Rapid testing closer to
the manufacturing line

22. What is the technology that you think will change biosafety testing of your manufactured
biopharmaceutical?
• Rapid microbiology
• Next generation sequencing
• Mass Spectroscopy
• Raman Spectroscopy
• Other rapid molecular techniques
• Enclosed manufacturing
Biosafety Testing Evolution | Adam Inche | 12March2020
Poll Question 2
22

23. Biosafety Testing Evolution | Adam Inche | 12March202023
Isoenzyme, 2015 – a lesson on change from the recent past
• Enzyme isoforms to identify
cell species
• Example test in ICH Q5A for
Cell Line Characterization
• Innovative Chemistry sole
supplier to industry
Dec ‘14 Innovative
Chemistry become
unresponsive
Feb ‘15 Customers
made aware of
supply issues
Mar ‘15 Customers notified
of Isoenzyme withdrawal –
alternative ID assay offered
Aug ‘15 Launch of
CO1 Barcode assay
Mar ’16 Pass x00
samples run on CO1
Barcode platform
201620152014
Internal quality systems remain
the biggest barrier to change
Large portion of ID testing
remains on DNA Fingerprint assay

24. adam.inche@milliporesigma.com
Adam Inche, DPhil
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