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B i o P r o c e s s TECHNICAL

Best Practices for Critical


Sterile Filter Operation
A Case Study

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Yanglin Mok, Lise Besnard, Terri Love, Guillaume Lesage, and Priyabrata Pattnaik

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A
number of regulatory

O
Figure 1:  Design and layout of Millidisk and Millipak barrier filters
guidelines recommend preuse
integrity testing of critical
sterilizing liquid filters for

N
aseptic processing (1–3). Before Support
Membrane
sterilization, a preuse test will confirm structure

IO
that a filter is installed properly and
was not damaged during shipment or
handling. Performing a preuse test after

S
Three One
sterilization detects damage that may hydrophilic hydrophobic
membranes

IS
have occurred during the sterilization membrane
cycle. Testing after sterilization limits One disk pair
risk, so it is a practice applied based on
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risk assessment. Because it is perceived
to reduce business loss risk, preuse post-
sterilization integrity testing (PUPSIT)
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is a current industry practice especially Gas


Liquid Air from integrity
in manufacturing products that will be
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Wetting and test and blow-


marketed in the European Union (EU). flushing fluids down can vent
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Unfortunately, it can be difficult to evacuate to drain from same filter


perform a PUPSIT without breaching
system sterility. A number of methods
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have been developed for running Outlet


PUPSIT and performing line
IT

conditioning without compromising


sterility. Such methods use a flush use the downstream hold tank as a sterilizing-grade Durapore membranes,
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bag, catch-can/flush bottle, or filter sterile boundary. both on the top and bottom of each
arrangement to create a sterile Here we describe a robust and disc (Figure 1). Because of that unique
boundary on the downstream side of versatile approach to PUPSIT using a combination of different membranes in
the product filter. Some applications self-venting, all-in-one sterile barrier parallel configuration, the devices can
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membrane filter from EMD filter condensate, steam, wetting liquid,


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Millipore, the life science business of and gases without compromising the
Product Focus:  All biologics Merck KGaA, Darmstadt, Germany, sterility of a steam-sterilized,
Process Focus:  Downstream which operates as MilliporeSigma in autoclaved, or gamma-irradiated
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processing, fill and finish


the United States and Canada. We system.
also include filtration line design Barrier filters can be used
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Who Should Read:  Process considerations for implementing downstream of sterilizing-grade filters
development and manufacturing barrier filters. to maintain system sterility. Water can
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pass through the hydrophilic discs


Keywords:  Filter integrity testing, Barrier Filters during a flushing sequence; air can
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aseptic processing, multiuse and


Millipak and Millidisk barrier filters pass through the hydrophobic discs
single-use technologies, barrier filters
are stacked-disc devices that combine during integrity-test and drying
Level:  Intermediate hydrophilic and hydrophobic sequences. A barrier filter acts as an

28 BioProcess International 14(5) M ay 2016


Figure 2:  Examples of sterile boundary designs Figure 3:  Barrier filters in a typical single
Catch Can filtration system (filling-line example)
Barrier Filters or Flush Bottle Flush Bag Downstream Filters Class C Class B/A
P P P
Integrity
Method tester
diagram Gas
Filter
Sterile Boundary Attribute

k ion
P P Filling line
Product

tanulat
(All maintain
sterility of
filter

rm
Fo
product filter
during preuse
test.) T T Liquid
Filter
Retest Barrier
filter
Ability to dry
the product
filter cycle ends, the low-point vent will be
Simple closed. The system then cools down
design with application of compressed gas (to
maintain positive pressure as well as
automatic vent during testing and and in filter drying after flushing to sterility in the filtrations system).
drying phases. The volume of particle- minimize product dilution. Wetting: The second step ensures
free water and air that can pass that a product filter is totally wetted
through these filters is unlimited. Filtration Line with particle-free water for its
Configuration and Operation integrity test. This step also flushes
Flushing and Testing For critical filter applications such as away extractable residues from a
Critical Product Filters final product-filling lines, barrier filters sterilized product filter element. To
Although filter rewetting and retesting can be used to provide a sterile boundary keep a filter train independent, the
should remain an optional activity while meeting regulatory requirements vent on the product filter is left open
when preparing a product final filter for preuse integrity testing. Typically, initially, and the isolation valve to
(sterilizing-grade filter) in line, the such filters are installed downstream of a downstream equipment is closed. The
PDA Technical Report 26 suggests up product filter (Figure 3). product filter vent is closed after air in
to three repetitions (3). The number of Figures 4 and 5 illustrate stepwise its housing has been vented. Water is
retests should be considered when use of barrier filters to provide a sterile directed to the drains through the
sizing for a flush bag. Barrier filters boundary in a stainless steel and a barrier filters. Isolating the flow path
could provide a more versatile solution. single-use filtration system with a to the barrier filters can enhance
Filters that are not wetted efficiently single product filter. Regulators wetting for increased applied pressure
the first time could give false failed test recommend redundant filtration as a drop through a product filter.
results. If rewetting volume is limited, risk-mitigation strategy for critical Testing: PUPSIT is either a
end users might discard integral filters filtration applications. Redundant diffusion or bubble-point test
that only marginally failed because of filtration is a type of serial filtration in (depending on the product filter). In all
improper wetting. Doing so could lead which a second product filter is used as cases, pressurized gas is applied on the
to unnecessary quality investigations as a back-up to protect against the product filter’s upstream side, which is
well as downtime associated with possibility of an integrity failure for the isolated from the system elements both
setting up a system again before use. primary product filter (3). Each filter upstream and downstream. Only the
Using a barrier filter allows for must be independently integrity- drain line with the barrier filters
rewetting and retesting with ease. testable in compliance with the relevant remains open to allow the free flow of
Unlimited volumes of particle-free regulations or guidelines. The step-by- test gas through the hydrophobic
water can be filtered through them to step approaches in Figures 4 and 5 can portion of their membranes.
wet these filters efficiently. Using a be applied to the second product filter Drying: Before product is
flush bag and/or catch-can instead in a redundant filtration system. introduced into the filtration line, the
creates a large footprint and could limit Sterilizing: The process begins with product filter typically is blown down
rewetting. Figure 2 summarizes the sterilization of a filtration system by and dried to prevent dilution of the
advantages and disadvantages of each SIP, autoclaving, or gamma-irradiation. product stream. The associated gas is
sterile boundary method available. If the chosen method of sterilization is vented through the barrier filter.
Barrier filters also allow SIP, then the barrier filter’s low-point Barrier Filter Integrity Testing:
extractables flushing through a sterile vent on its upstream side will be kept Millipak and Millidisk barrier filters
boundary to drain. A barrier filter also open during the SIP cycle. Condensate, are integrity tested offline using 70/30
can be used as a vent in system cooling steam, and air will pass through the isopropyl alcohol (IPA) as the wetting
after steam-in-place (SIP) sterilization filter to drain on its outlet. When the fluid.

30 BioProcess International 14(5) M ay 2016


Process: Once the integrity of the straightforward (Figures 4 and 5). flushing or wetting liquid passes
product filter is confirmed, the sterile Similar to all critical applications, though the barrier filters to a drain.
filtration process can begin. important process steps and conditions Flushing/wetting conditions are derived
After Processing: After the sterile should be reviewed during system and from vendor recommendations (5, 6) and
filtration process, product recovery process design to ensure successful can vary among product filters. Inlet
through a sterilizing-grade filter can implementation of this application. pressures on barrier filters during
be achieved through air blow-down Verification testing should be performed flushing/wetting procedures should not
with application of a low differential before implementation of an assembly exceed 0.7 bar. If enhanced wetting of a
pressure (air or nitrogen) of 5 psi to with barrier filters for product filtration. product filter is required, higher static-
the filter. Users can apply a buffer Sterilization of Filtration System: hold pressure can be implemented
chase, but product dilution must be Sterilization renders a system or across it. However, the downstream
accounted for. At the end of product equipment free of microorganisms and section of that product filter (including
recovery, sterilizing-grade filters are is a critical step, especially for aseptic the barrier filter) should be isolated
integrity tested with particle-free, manufacturing processes. Filters can be during such a high-pressure hold step.
water-based, alcohol (70/30 IPA/ sterilized through SIP for Millidisk Key Verification Point — Efficacy of
water), or product-based integrity test format (cartridge filters) and Product-Filter Wetting: The efficiency
specifications. For particle-free–water- autoclaving or gamma irradiation for of wetting a product filter(s) through
based or alcohol integrity-test Millipak (disposable capsule) filters barrier filters can be verified by
specifications, a filter must be flushed (Figure 6). Thermocouples, radiation performing product filter integrity
adequately with the test liquid to dosimeters, and biological indicators testing. Results can be compared with
remove residual product before testing. serve as the worst-case positions within filter specifications and past trending.
Product-based integrity-test a filtration system and assembly for Key Verification Point — Gas Flow
specifications can be developed validation of sterilization. Rate of Barrier Filters After Flushing/
through support from filter vendors. Flushing and Wetting Product Wetting Procedure: Ensuring that the
Filters: A filtration system may be hydrophobic membrane in a barrier
Design Considerations flushed and wetted to remove filter remains dry is critical. Such
Use of Millipak and Millidisk barrier extractables after sterilization as well as dryness can be verified in the filter
filters in a filtration line is simple and for product filter integrity testing. The following a flushing/wetting procedure

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during the qualification phase with a their gas flow-rate level at 100 mbar pressure for five minutes. Considering
low-pressure bubble-stream test. This (1.5 psi) pressure, then comparing that the average hydrophobicity level of
includes disconnecting barrier filters to the nominal gas flow-rate level of a polyvinylidene fluoride (PVDF) discs,
from their assembly and determining new filter unit wetted optimally at low flow rates in the 30–100% range of
nominal rate are characteristic of a
Figure 4:  Product filter preparation steps for steamable line, from sterilizing to integrity testing “breathing” unit.
Key Verification Point — Using
Product filter 1 Sterilizing 2 Wetting 3 Testing Wetting Medium Apart from Water for
preparation Integrity test Injection (WFI): Compatibility and
steps for Close valve of product

steamable line: once wetted. filter intrusion pressure/wettability for the


from sterilizing hydrophobic filters within barrier
to integrity Product
filters should be verified before the
testing Steam
in place
filter
Atmospheric
filters are used. Wetting hydrophobic
Water
filters can reduce their air-flow
Barrier (max. capacity, leading to increased pressure
Valves filter 0.5 bar)
drop across the filter assembly during
Open
integrity testing or product-filter
Closed Vent steam through Drain water through Vent test gas
barrier filter. barrier filter. through barrier filter. blow-down. Millipak and Millidisk
barrier filters validation guides provide
4 Drying 5 Barrier Filter 6 Processing Product filter chemical compatibility information
Compressed air
Testing preparation summaries (7–9).
Close steps for
valve
once steamable line: Integrity Testing
wetted.
from drying
of Product Filter
to processing
Filtration assembly designers should
70%/30%
include strategies to minimize product
IPA/water Product hold by reducing piping or tubing
Barrier filter Valves length and to ensure maximum product
tested off-line
Open recovery. The strategy for integrity
Vent air through Closed testing product filters also should be
barrier filter. well thought-out, especially for
redundant filtration assemblies. In
Figure 5:  Product filter preparation steps for single-use assemblies, from sterilizing to integrity 2012, Felo and coworkers at
testing and from drying to process MilliporeSigma provided an in-depth
Product filter look at how product filters can be
1 Sterilizing 2 Wetting 3 Testing integrity tested in a single-use assembly
preparation Water
steps for (max. 0.5 bar) (10). Their strategy can be applied to
single-use Close valve stainless steel systems as well.
Air filter once wetted.
assemblies: Interference on Product Integrity
from sterilizing Testing from Barrier Filters: Barrier
Product
to integrity Product
filter filter Product
filters are placed downstream of a
filter
testing
product filter. To verify the absence of
Valves Barrier interference, the integrity test result of
filter
Open the product filter both with and
Sterilized by autoclave
or gamma irradiated as Drain water through Vent test gas without the barrier filters can be
Closed part of an assembly barrier filter. through barrier filter.
compared. Those results should fall
within 70 mbar for a bubble-point test
4 Drying 5 Barrier Filter 6 Processing Product filter and 5% for diffusion flow.
Testing
Compressed air

preparation Failure Mode Test: To simulate a


Product
steps for worst-case scenario (failure-mode
Close valve
once wetted. single-use test), users can examine how a fully
assemblies:
70%/30% wet barrier filter gas flow rate
IPA/Water from drying
to processing
compromises a product-filter integrity
test. Millipak and Millidisk barrier
Barrier filter filters can be fully wetted by flushing
tested off-line
Valves with WFI at 3 bar.
Open Adaptation of Troubleshooting
Vent air through
barrier filter.
Closed Decision Tree: If a product filter fails
its integrity test, users can apply a
32 BioProcess International 14(5) M ay 2016
Figure 6:  Sterilization considerations for Millidisk and Millipak barrier filters
troubleshooting decision tree such as
the example given in PDA’s Technical Millidisk Barrier Format Millipak Barrier Format
Report #26 (3).

Filter Format
Drying of Filtration System
To minimize product dilution or
contact of product with the wetting
liquid (either buffer or water) before
filtration, the assembly may be blown
down to remove wetting liquid. The
current industry practice of blowing Steam-in-Place Autoclave

Important Sterilization
down a filtration system ranges from Sterilization process validation required

Considerations
30 minutes to three hours. Maximum 135 °C for 60 minutes, Maximum 123 °C for 90 minutes,
Duration of Drying: Exact drying up to four times up to three times
times should be verified on site and Steamed in forward direction with 5 psi Has no vent (self-venting) and serves as
(340 mbar) maximum pressure drop the sterile vent for air and steam of the
determined during qualification by
Housing should a include housing drain autoclaved assembly
weight and visual checks. The same for condensate removal.
time taken to reach the “dry weight”
of the assembly will be required for
drying the assembly during operation. barrier filter should be integrity tested 5 P35515 Rev G: Wetting Instructions for
Filter Units with Durapore Membrane. EMD
Acceptable Applied Pressures: after the product filter has passed
Millipore: Billerica, MA, April 2012.
Typical pressures applied for drying integrity but before product filtration.
6 RF1510EN00: Hydrophilic Durapore
filtration assemblies are 0.5 bar higher This minimizes the risk of Cartridges and Capsules User Guide. EMD
than the bubble-point pressure of a reprocessing product because of a poor Millipore: Billerica, MA, January 2002.
product filter. Such pressures should installation or nonintegral barrier 7 VG026 rev 2: Millidisk Cartridge Filter
not exceed the maximum allowable filter caused by mishandling. For Units with Hydrophilic Durapore Membrane
pressure of the “weakest link” in an situations in which resources are Validation Guide. EMD Millipore: December
1999.
assembly. That might be silicone limited and product can be
8 VG033 Rev D: Millipak Disposable Filter
tubing, a connector, or a barrier filter reprocessed, barrier filters can be
Units Validation Guide. EMD Millipore:
(4.1 bar for Millipak and Millidisk integrity tested after product filtration. Billerica, MA, May 2012.
formats), for example. If the required 9 VG2000EN00: Millidisk Barrier Filter
pressure is greater than what the For Best Practices Validation Guide. EMD Millipore: Billerica,
weakest link allows, then blow-down Barrier filters help enable best MA, May 2002.
pressure should be reduced, and an practices of aseptic filtration lines for 10 Felo M, Oulundsen G, Patil R. Single-

2012: 38–41. •
extended drying time can be applied. flushing/wetting and preuse integrity Use Redundant Filtration. BioPharm Int. 25(4)
Absence of Air-Flow Interference: testing of product filters. In particular,
Restriction of air flow through fittings, implementation of Millipak and
connectors, tubing, or piping used in an Millidisk barrier filters is easy and Corresponding author Yanglin Mok, BE, is a
senior process engineer and technical
assembly should be minimized. As a provides flexibility and versatility to
manager of the Biomanufacturing Sciences
product filter dries, the air-flow rate the filtration line.
Network, 1 Science Park Road, #02-10/11 The
will increase and pressure drop across Capricorn, Singapore 117528; 65-6403-5313,
the product filter will decrease. References fax 65-6403-5322; yanglin.mok@
1 Annex 1: Manufacture of Sterile
merckgroup.com. Lise Besnard, MSc, is a
Medicinal Products. Volume 4, EU Guidelines to
Integrity Testing process development scientist at Sanofi
Good Manufacturing Practice Medicinal Products
of Barrier Filters for Human and Veterinary Use. European Pasteur, 1541 Avenue Marcel Mérieux, 69280
Barrier filters are integrity tested Commission: Brussels, Belgium, November Marcy l’Etoile, France. Terri Love, BSc, is a
offline with 70/30 IPA/water as a 2008; http://ec.europa.eu/health/files/eudralex/ biomanufacturing engineer; Guillaume
wetting medium and bubble-point test vol-4/2008_11_25_gmp-an1_en.pdf. Lesage, MSc, is a biosafety technical
specification of ≥1,280 mbar 2 CBER/CDER/ORA. Sterile Drug Products consultant; and Priyabrata Pattnaik, PhD,
Produced By Aseptic Processing: Current Good is director of the worldwide vaccine initiative;
(18.5 psi). These filters can be wetted
Manufacturing Practice. US Food and Drug all with the life-science business of Merck
by dynamic flushing or static-soak Administration: Rockville, MD, September 2004. KGaA, Darmstadt, Germany, which operates
methods. The wetting procedure of 3 Technical Report No. 26: Sterilizing as MilliporeSigma in the United States and
barrier filters can be found in a Filtration of Liquids. Parenteral Drug Canada. Millipak, Millidisk, and Durapore
technical guide (5). A 15-minute static Association: Bethesda, MD, 2008. are registered trademarks of MilliporeSigma.
soak can be applied to either Millipak 4 ASTM Standard F838-83: Standard
or Millidisk barrier filters. Test Method for Determining Bacterial Retention
For reprints, contact Rhonda Brown of Foster
of Membrane Filters Utilized for Liquid
For critical product applications in Printing Service, rhondab@fosterprinting.com,
Filtration. American Society for Testing and
which resources are readily available, a Materials: Philadelphia, PA, 1983.
1-866-879-9144 x194.

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