Sampling in an Aseptic Process
Risk Mitigation & Regulatory Compliance
Somasundaram G.
Senior Technical Consultant, Asia Pacific
Process Solution, Merck KGaA
September 23, 2020
2
Agenda
1
3
4
Introduction to Sampling – Where, What & Challenges
Key considerations for Closed & Disposable sampling options
2 Regulatory recommendations and corresponding needs – Key Drivers
Complexity and Risks of traditional sampling methods
3
Introduction to Sampling
Where, What & Challenges
4
4
– Representative Sample :
– Composition
– Non-operator dependent
– Repeatable
– Protect the Process (Contamination-free)
– Protect the Person (Contained)
– Protect the Product (Contained)
– Easy of Use
– Save Time and Cost Efficient
Introduction to sampling
What are The Requirements and Challenges?
Process
People Sample
Validated
Sampling
Procedure
Validated
Sampling
Procedure
Equipment
In today’s biopharmaceutical market, sampling is critical during every step in the manufacturing process. An
imprecise or false positive result can lead to a quarantine as well as the need to repeat the analysis.
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Sampling plan & options
USP BioreactorCell culture
media
Cell culture
supplement
Media
filtrationClarification
BBR &
Intermediate
pool
Broad
considerations
• Bioburden levels of appropriate batches to be tested based on
statistical/risk assessment
• Inoculum expansion to first active control expansion step (at scale):
Bioburden needed
• Good practice to retain samples at each expansion operation until batch
release
If clarified harvest
is filtered into a
pooling tank.
Recommended to
test bioburden of
a few batches of
clarified harvest
before filtration
Typically
prefiltration
samples (entry
point) or after
hold times
(proliferation)
Each protein pool
should be tested
for bioburden &
endotoxin
Via routine sampling
during bioreactor run
• Routine sampling may not be needed when max
preparation & filtration durations are established
• Periodic monitoring recommended annually/after
prolonged shutdowns
What are
typically
sampled?
• Cell Viability (count)
• Bioburden, including
for continuous cell
lines
• Endotoxin (not at
inoculum expansion)
• Purity check
• Osmolality
• Metabolite
• Bioburden
• Endotoxin
• pH
• Conductivity
• Media Concentration
Endotoxin
Bioburden
TOC
• Endotoxin
• Bioburden,
including
pooled
samples (prior
to filtration)
Where to Sample
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Sampling plan & options
DSP1 AffinityVirus
Inactivation
Aggregate
RemovalCEX AEX
Virus
Filtration
Broad
considerations
• Bioburden control: some of the unit operations may not be a closed operations / or
sterilizable
• Bioburden & endotoxin testing: WFI rinse (retentate/drain line), WFI rinse (after pre-use
sanitization), EQ buffer (after equilibration step), WFI rinse (after post-use cleaning)
• Establish microbial control of reusable items (e.g. filters, resins). Acceptance
specifications for new & unused resins depends on vendor (typically ≤100 CFU/mL)
What are
typically
sampled?
• Endotoxin
• Bioburden
• TOC
• Conductivity
• Osmolality
• Impurities
• Osmolality
• pH
• Conductivity
• Bioburden
• Endotoxin
• Impurities
• Virus
inactivation
kinetics?
• Impurities?
• Endotoxin?
• Endotoxin
• Bioburden
• TOC
• Conductivity
• Osmolality
• Impurities
• Osmolality
• Endotoxin
• Bioburden
• TOC
• Conductivity
• Osmolality
• Impurities
• Osmolality
• pH
• Conductivity
• Bioburden
• Endotoxin
• Impurities
Where to Sample
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Sampling plan & options
DSP2 /
Prep zoneUFDF
Sterile
Filtration
Sterile Bulk/
Dispensing
Operations
Buffer MixingBuffer
Holding
Buffer
Filtration
Broad
considerations
• Bioburden control: some
of the unit operations
may not be a closed
operations / or
sterilizable
• Product purity, TOC
• pH control
• Via routine sampling
• Many configurations.
• Sampling should take
risk associated with
sample & nature of
filling operation, into
consideration
• Bioburden levels of appropriate batches to be
tested based on statistical/risk assessment
• Final DF/formulation buffers: Endotoxin testing
recommended prior to use for all batches.
• Non-filtered buffer: Recommended to be
prepared fresh/used ASAP. For >24h: Hold
time validation (bioburden & endotoxin)
needed.
What are
typically
sampled?
• Endotoxin
• Bioburden
• TOC
• CIP
validation
• Impurities
• Osmolality
• Biobur
den
• Endotoxin
• Bioburden
• pH
• Conductivity
• Buffer conc
• pH
• Conductivity
• Endotoxin
• Bioburden
• Buffer conc
Where to Sample
Aseptic Filler
Sterile Hold Tank
Vent FilterVent Filter
Bioburden
Reduction
Filter
Sterilizing
Filter
Sterilizing
Filter
Prefiltered
Formulation
Sampling pointFinal Filling
Single SGF
Sampling plan & options Where to Sample
Aseptic Filler
Sterile Hold Tank
Vent FilterVent Filter
Bioburden
Reduction
Filter
Sterilizing
Filter
Sterilizing
Filter
Vent Filter
Sterilizing
Filter
Prefiltered
Formulation
Redundant Filtration
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How to ensure that the process is microbiologically
controlled?
• Regular sampling of equipment that can not be sterilized
• Collection of samples after cleaning and sanitization
• Collection of samples during equipment equilibration and rinsing
– BPOG (2015):
– “...containers include disposable systems that are closed to the environment“
– “Bioburden samples are recommended to be stored at 2-8oC and tested within 24 hours
of collection“
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Setting bioburden control levels (Alert/ Action)
1
0
– BPOG (2015):
– “There are no recommended bioburden levels provided in regulatory guidelines or
compendia... Manufacturers are responsible for setting bioburden control levels...“
• Most members: 1 to 10 CFU/mL. Upstream (conducive for proliferation): Near sterile;
Downstream: ≤ 10 CFU/mL
• Control levels are set prior to product licensure. “After commercialization, periodic
review is recommended to ensure control levels reflect product performance over
time.“
• Correlate bioburden testing with results of routine environmental monitoring.
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Regulatory recommendations and
corresponding needs –
Key Drivers
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Drivers for Aseptic Process Sampling
Assess the state
of the process
• Adjust sensors
• Verify, detect,adjust parameters
Transfer materials
• Adjustment by addition
• Seeding by inoculation
Extract materials for
later assessment
• Internal investigation
• Regulatory requirements
• Secure traceability
Global
Regulatory
trends:
• Process Validation
• QbD
• PAT
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Relevant Regulatory Agencies & Industry Associations
Regulatory Agencies /
Industry Associations
Relevant
Documentation
Food and Drug Administration cGMP guidance
World Health Organization
Annex 2 – Good manufacturing guidance for API
Annex 4 - Good Manufacturing Practices for pharmaceutical
products: main principles
European Medicines Agency GMP Annex 1 - Manufacture of Sterile Medicinal Products
International Council for Harmonization of Technical
Requirements for Pharmaceuticals for Human Use ICH Q7 – GMP guidance for API
Parenteral Drug AssociationReport #69 - Bioburden and Biofilm Management in
Pharmaceutical Operations
The Pharmaceutical Inspection Convention and
Pharmaceutical Inspection Co-operation Scheme
Guide to good manufacturing practice for medicinal products –
Part I
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Extracts of Relevant Requirements
Recommendations Source
Contamination control&
Monitoring before bioburden reduction
• PICS/S – FDA cGMP – Part 1 §5.19 –f • Use of closed system recommended from phase 1
• WHO Annex 4• “The use of disposable sampling materials has distinct advantages”
• WHO Annex 2 - ICH Q7A - GMP guidance for API• EU GMP Annex 1 • EudraLex – Vol 4 – Part II – 2009
Operator bias elimination• WHO Annex 4• FDA • European Pharmacopeia - Guidelines for Sampling of Pharmaceutical
Products and Related Materials
Representative sample
Health & safety focus
Retained samples
• FDA cGMP Guidance for the industry investigational drugs section F. Laboratory Controls / 1. Testing
• 2 years after expiration date / completion of trial and twice the quantity necessary to perform all tests
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Sampling Process & Precautions
WHO Annex 4: Preparation for
sampling
• “All sampling tools and
implements should be made
of inert materials and kept
scrupulously clean. […].”
• “The cleaning procedure
used for all sampling tools
and implements should be
documented and recorded.”
• “The use of disposable
sampling material has distinct
advantages”
WHO Annex 4: Sampling
Operations and Precaution
“There should be a written
procedure describing the
sampling operation. […]. It
should ensure that
representative samples are
taken in sufficient quantity for
testing in accordance with
specifications.”
WHO Annex 4: Storage and
Retention
“The container used to store a
sample should not interact with
the sampled material nor allow
contamination. […]. As a general
rule the container should be
sealed and preferably tamper-
evident.”
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Sampling of Pharmaceutical Products and
Related Materials
• FDA cGMP for phase 1 drugs
“recommends the use of closed system to minimize the risk of contamination”
•
Guidance for the industry investigational drugs section
F. Laboratory Controls / 1. Testing
“We recommend that the sample consist of a quantity adequate to perform additional testing or
investigation if required at a later date […]. We recommend that you appropriately store and
retain the samples for at least two years […].”
• Q7A GMP guidance for manufacturing API – section C. In-process Sampling and Controls (8.3)
“In-process sampling should be conducted using procedures designed to prevent contamination
[…]. […] ensure the integrity of samples after collection.”
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Regulatory Focus on Contamination
Control during Sampling
8. Production and in-process controls
• 8.3 In-process sampling and controls
• 8.35 “In-process sampling should be
conducted using procedures designed to
prevent contamination of the sampled
material and other intermediates or APIs.
Procedures should be established to
ensure the integrity of samples after
collection.”
Annex 2, WHO GMP for active pharmaceutical
ingredients
EudraLex, Volume 4
ICH Q7A
Identical script in these 3 regulatory documents
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8.94 Bioburden samples should be taken from the bulk product and
immediately prior to the finalsterile filtration. Systems for taking samples should be designed so as not to introduce contamination.
9.7 Sampling methods should not pose a risk of contamination to the manufacturing operations.
7.3 Non-essential processes such as product inspection
and in process testing should be conducted à with the crimping option
there is a validated and easy disconnection in place to further
treat the sampleoutside the clean areas wherever
possible.
Annex 1 EU GMP (Draft)Regulatory requirements
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Complexity & Risk of
Traditional Sampling Methods
2020
Open Sampling Valve
Steam in place valve
Septum sampling Aseptic Connectors Tube Welding
Large number of
samples
Low cost per sample
Large number of samples
Closed sampling
Low cost per sample
Large number of
samples
Low cost per sample
Flexible & Reliable
Aseptic sampling
Safe and disposable
Large number of samples
Aseptic sampling
Flexible
Dead-leg
Loss of product (flush)
Open sampling
Impossible to sterilize
Complex operation (SIP)
Safety hazard (heat)
Risk of sample dilution
Container limitations
Not steam sterilizable
Safety hazard (needles)
Sample volume
limitation
Extra cost
Potential dead-leg
(tubing)
Limited disconnection
Waste of product
Requires utility
Piece of hardware
High risk of
contamination
Operator & Process
safety
Sample
representativeness
Operator training & safety
Sample
representativeness
High risk of
contamination
Operator safety
Sample
representativeness
Operator trainingRequires maintenance
Operator training
What are the Ways to Sample?Traditional sampling: Pros, Cons and Limitations
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Complexity and risks of traditional sampling methods
• FDA Warning letters (483) issued in
2015
• 104 – Procedures designed to prevent
microbiological contamination of sterile
drug products not established, written
or followed
• 24 – Representative samples not
obtained
Microbial contaminations during manufacturing
• Increases risk
• Results in enormous cost
• Requires complicated investigations to
prevent reoccurrence
Cost of sampling VERSUS cost of a shut down
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30Percent
1-6Months
1-14Million Euro
Percent of process deviations
caused by contamination*
Length of time to complete an investigation
Operations cost
*Source Langer 2013, Wiebe 2014
Biologics in-process contamination
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30Percent
1-6Months
1-14Million Euro
*Source: Langer 2013, Wiebe 2014
Biologics in-process contamination
Impact Productivity losses
Material replacement costs
Batch loss
Interruption of product supply
Delay in clinical development
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Key considerations for Closed
& Disposable sampling
options
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Considerations for adopting single-use
Aseptic sampling
Situations Implications
Contamination of samples
(out of in process
specifications)
• Product batch on-hold
• Internal investigations
• Additional tests on final product
• Conflict between manufacturing and QAQC
• Risk of remarks from regulatory inspectorsContamination of process
Complex sampling procedure
• Risk of false results
• Frequent operator training
• Unhappy operators
• Internal investigations
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Considerations for adopting single-use
Aseptic sampling
Indirect
cost
Direct
cost
• Labor costs
• Process/ product
downtime
• Scrap of batch due to
contamination
• Investigations for
product/ process
deviations
• Purchase price
• Taxes
• Freight
• Inventory costs
Total cost of
ownership
(TCO)
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Self-assessment: Am I suitable to adopt single-use
Aseptic sampling?
Considerations Considerations
1 2 3 1 2 3
It is difficult to implement changes to
my infrastructure (e.g. piping).
There is high risk of bio-
contamination in my process.
There are challenges to conduct
cleaning validation.
My current sampling method has
risk of cross-contamination.
My operators are very familiar with
plastic components in the process.
I need to collect samples anywhere.
I have limited process time to
prepare sampling assemblies
(traditional).
I need to collect large numbers of
samples.
I have limited cycle time. I need
quick turnaround between batches.
I require a varied range of sampling
containers.
I am concerned about operator
safety.
My process is sensitive to price.
I have limited resources for operator
training.
Strong
YES
Strong
NOStrong
YES
Strong
NO
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Sterile sampling value assessmentCase study example
62% Reduced labor hours
35% Reduced product loss during
sampling
80% Reduced deviation costs
400 Autoclave cycles eliminated10%Overall cost reduction
Key facts
Background
Replace standard open sampling with NovaSeptum® sterile sampling assembly
Outcome
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• There are complexity and heightened risks in using traditional
sampling methods.
• Regulatory recommendations are in place and single-use aseptic
sampling could help meet several corresponding needs.
• There are several key considerations to design/optimize your sampling
plan.
Conclusions
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Acknowledgements
Mark Antoine Kaag, Global Product Manager, France
Janmeet Anant, Global Regulatory Advocate, USA
Thank You