Bioburden Control at

the Sterile Filtration

Step: A risk-based
approach
Presenter: Ray Field, Director, Biopharmaceutical
Development & CMC TL
Co-authors: Harry Yang, Sr. Director, Biostatistics;
Derek Murphy, Director, QA; John Alvino, Manager,
Regulatory Affairs; Stephen Chang, Assoc. Director,
Biopharmaceutical Development
06 May 2013 presentation to EBE
Manufacture of a Sterile Drug Product

 Microbial control during manufacturing is critical for ensuring product

quality and safety.
 Sterile biologic drug products (finished dosage forms) are typically
manufactured by sterile filtration followed by aseptic filling and
processing.
 Control of microbial load at the sterile filtration step is an essential
and required component of the overall microbial control strategy.
Drug Substance Receipt at Typical
Fill Facility and Storage Biologics
at 2-8°C Manufacturing
Pre-Filtration Flow Diagram
↓ Bioburden Test
for a Liquid
Sterile Drug
Sterile Filtration Product

↓
 In-process control
Aseptic Filling and bioburden sample
Stoppering pulled immediately
prior to sterile
↓ filtration step

100% Visual Inspection  Process assumes

no pooling, mixing,
or compounding
↓
Drug Product Storage
at 2-8°C

3
Regulatory Guidance
EMA/CHMP/BWP/534898/2008
 Maximum bioburden to be stated in control of sterile filtration step
 Establishes typically accepted limits for number of CFU and sample
test volume
 Allows exceptions, with use of a pre-filter or, if justified, for limited
product availability (but does not define “limited”)

– Excerpt from EMA guidance:

“For sterilisation by filtration the maximum acceptable bioburden prior to
the filtration must be stated in the application. In most situations NMT
10 CFU/100 ml will be acceptable, depending on the volume to be
filtered in relation to the diameter of the filter. If this requirement is not
met, it is necessary to use a pre-filtration through a bacteria-retaining
filter to obtain a sufficiently low bioburden. Due to limited availability of
the formulated medicinal product, a pre-filtration volume of less than
100 ml may be tested if justified.”
Modeling Bioburden Distribution in Solution

 Negative binomial (NB)

distribution can be used to
model distribution of bacteria.
 NB model allows for bacterial
clumping with better
representation of
“microbiological environment”
compared to uniform (Poisson)
distribution.
 Statistical analysis shows
sensitivity (probability) to detect
10 CFU/100 mL = 41.2%.
 At same 41.2% sensitivity level,
corresponding bioburden level
for different sample test
volumes can be calculated.
Alternate Test Volumes at 41.2% Sensitivity Level
 Smaller test volumes means that higher bioburden levels are
needed to give equal probability to detect.
– For example, equal probability to detect bioburden level of 16.2/100 in
10 mL as 10/100 in 100 mL

Test Volume (mL) CFU/100mL

10 16.2
20 12.6
30 11.4
40 10.9
50 10.6
60 10.4
70 10.2
80 10.1
90 10.1
100 10

6
Employ risk-based approach to justify alternate
testing schemes

 There is a risk associated with the limited sensitivity of bioburden

test method:
Risk = Severity x Probability of failing to detect bioburden
 Opportunity to apply a risk-based approach for pre-filtration
bioburden testing:
Quantitate risk of failure to detect bioburden for a given bioburden
testing scheme (test limit and sample volume)

Reduce risk to within acceptable risk tolerance level through process

capabilities/design and overall microbial control strategy (see next slide)

Refine bioburden testing scheme, if needed, and justify pre-filtration

bioburden test limit and sample volume based on risk assessment
results and control strategy
Some potential process design and control
strategy risk mitigation measures
 Reduce microbial load prior to and on sterile filter
– Remove bioburden with upstream filtrations and/or pre-filters
– Limit hold times and room temperature storage
– Implement aseptic handling techniques where appropriate
 Improve capability of sterile filtration step to prevent bioburden
breakthrough
– Select and validate sterile filter membranes with high microbial retention
capabilities (e.g., ≥ 107 CFU/cm2)
– Increase effective filter surface area by using larger single filter or
multiple filters in series
– Limit batch volume to be sterile filtered
– Test integrity of sterilising filters pre- and post-use
 Test and control bioburden throughout manufacturing process,
including during raw material introduction, to reduce bioburden
breakthrough risk across sterile filtration step
Conclusions

 Pre-filtration bioburden control should be considered one important

component of the overall microbial control strategy for the entire
manufacturing process.
 Alternative bioburden test limits and potentially smaller sample
volumes compared to 10 CFU/100 mL could be justified through a
risk-based approach that considers process capabilities and overall
control strategy.
 Such risk-based approaches are consistent with current regulatory
guidance and expectations.
Acknowledgements

 Steve Bishop, Director, Biopharmaceutical Development

 Gail Wasserman, Sr. Vice President, Biopharmaceutical
Development
 Ann Warford, Orit Scharf, Leslie Day, Pat Cash, Claudia van
Elderen, and Michael Li