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Best Practices for Critical Sterile Filter Operation

A Case Study

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

A number of regulatory

guidelines recommend preuse

integrity testing of critical

sterilizing liquid filters for

aseptic processing (1–3). Before

sterilization, a preuse test will confirm

that a filter is installed properly and

was not damaged during shipment or

handling. Performing a preuse test after

sterilization detects damage that may

have occurred during the sterilization

cycle. Testing after sterilization limits

risk, so it is a practice applied based on

risk assessment. Because it is perceived

to reduce business loss risk, preuse post-

sterilization integrity testing (PUPSIT)

is a current industry practice especially

in manufacturing products that will be

marketed in the European Union (EU).

Unfortunately, it can be difficult to

perform a PUPSIT without breaching

system sterility. A number of methods

have been developed for running

PUPSIT and performing line

conditioning without compromising

sterility. Such methods use a flush

bag, catch-can/flush bottle, or filter

arrangement to create a sterile

boundary on the downstream side of

the product filter. Some applications

Product Focus: All biologics

Process Focus: DownstreAm

processing, fill AnD finish

Who should read: process

Development AnD mAnufActuring

KeyWords: filter integrity testing,

Aseptic processing, multiuse AnD

single-use technologies, bArrier filters

level: intermeDiAte

28 BioProcess International

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May 2016

Figure 1: Design and layout of Millidisk and Millipak barrier filters Support Membrane structure Three
Figure 1: Design and layout of Millidisk and Millipak barrier filters
Support
Membrane
structure
Three
One
hydrophilic
hydrophobic
membranes
membrane
One disk pair
Gas
Liquid
Air from integrity
Wetting and
test and blow-
ushing uids
down can vent
evacuate to drain
from same lter
Outlet

use the downstream hold tank as a

sterile boundary.

Here we describe a robust and

versatile approach to PUPSIT using a

self-venting, all-in-one sterile barrier

membrane filter from EMD

Millipore, the life science business of

Merck KGaA, Darmstadt, Germany,

which operates as MilliporeSigma in

the United States and Canada. We

also include filtration line design

considerations for implementing

barrier filters.

Barrier Filters

Millipak and Millidisk barrier filters

are stacked-disc devices that combine hydrophilic and hydrophobic

sterilizing-grade Durapore membranes,

both on the top and bottom of each

disc (Figure 1). Because of that unique

combination of different membranes in

parallel configuration, the devices can

filter condensate, steam, wetting liquid,

and gases without compromising the

sterility of a steam-sterilized,

autoclaved, or gamma-irradiated

system.

Barrier filters can be used

downstream of sterilizing-grade filters

to maintain system sterility. Water can

pass through the hydrophilic discs

during a flushing sequence; air can

pass through the hydrophobic discs

during integrity-test and drying sequences. A barrier filter acts as an

Figure 2: Examples of sterile boundary designs

Barrier Filters Catch Can or Flush Bottle Flush Bag Downstream Filters Method P P P
Barrier Filters
Catch Can
or Flush Bottle
Flush Bag
Downstream Filters
Method
P
P
P
Gas
diagram
Filter
P
P
(All maintain
sterility of
product lter
during preuse
test.)
T
T
Liquid
Filter
Retest
Ability to dry
the product
lter
Simple
design
Sterile Boundary Attribute

automatic vent during testing and drying phases. The volume of particle- free water and air that can pass through these filters is unlimited.

Flushing and testing critical Product Filters

Although filter rewetting and retesting should remain an optional activity when preparing a product final filter (sterilizing-grade filter) in line, the PDA Technical Report 26 suggests up to three repetitions (3). The number of retests should be considered when sizing for a flush bag. Barrier filters could provide a more versatile solution. Filters that are not wetted efficiently the first time could give false failed test results. If rewetting volume is limited, end users might discard integral filters that only marginally failed because of improper wetting. Doing so could lead to unnecessary quality investigations as well as downtime associated with setting up a system again before use. Using a barrier filter allows for rewetting and retesting with ease. Unlimited volumes of particle-free water can be filtered through them to wet these filters efficiently. Using a flush bag and/or catch-can instead creates a large footprint and could limit rewetting. Figure 2 summarizes the advantages and disadvantages of each sterile boundary method available. Barrier filters also allow extractables flushing through a sterile boundary to drain. A barrier filter also can be used as a vent in system cooling after steam-in-place (SIP) sterilization

30 BioProcess International

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May 2016

and in filter drying after flushing to minimize product dilution.

Filtration line conFiguration and oPeration

For critical filter applications such as final product-filling lines, barrier filters can be used to provide a sterile boundary while meeting regulatory requirements for preuse integrity testing. Typically, such filters are installed downstream of a product filter (Figure 3). Figures 4 and 5 illustrate stepwise use of barrier filters to provide a sterile boundary in a stainless steel and a single-use filtration system with a single product filter. Regulators recommend redundant filtration as a risk-mitigation strategy for critical filtration applications. Redundant filtration is a type of serial filtration in which a second product filter is used as a back-up to protect against the possibility of an integrity failure for the primary product filter (3). Each filter must be independently integrity- testable in compliance with the relevant regulations or guidelines. The step-by- step approaches in Figures 4 and 5 can be applied to the second product filter in a redundant filtration system. Sterilizing: The process begins with sterilization of a filtration system by SIP, autoclaving, or gamma-irradiation. If the chosen method of sterilization is SIP, then the barrier filter’s low-point vent on its upstream side will be kept open during the SIP cycle. Condensate, steam, and air will pass through the filter to drain on its outlet. When the

Figure 3: Barrier filters in a typical single filtration system (filling-line example)

Class C Class B/A Integrity tester Product Filling line lter Barrier lter Formulation tank
Class C
Class B/A
Integrity
tester
Product
Filling line
lter
Barrier
lter
Formulation
tank

cycle ends, the low-point vent will be closed. The system then cools down with application of compressed gas (to maintain positive pressure as well as sterility in the filtrations system). Wetting: The second step ensures that a product filter is totally wetted with particle-free water for its integrity test. This step also flushes away extractable residues from a sterilized product filter element. To keep a filter train independent, the vent on the product filter is left open initially, and the isolation valve to downstream equipment is closed. The product filter vent is closed after air in its housing has been vented. Water is directed to the drains through the barrier filters. Isolating the flow path to the barrier filters can enhance wetting for increased applied pressure drop through a product filter. Testing: PUPSIT is either a diffusion or bubble-point test (depending on the product filter). In all cases, pressurized gas is applied on the product filter’s upstream side, which is isolated from the system elements both upstream and downstream. Only the drain line with the barrier filters remains open to allow the free flow of test gas through the hydrophobic portion of their membranes. Drying: Before product is introduced into the filtration line, the product filter typically is blown down and dried to prevent dilution of the product stream. The associated gas is vented through the barrier filter.

Barrier Filter Integrity Testing:

Millipak and Millidisk barrier filters are integrity tested offline using 70/30 isopropyl alcohol (IPA) as the wetting fluid.

Process: Once the integrity of the product filter is confirmed, the sterile filtration process can begin.

After Processing: After the sterile

filtration process, product recovery through a sterilizing-grade filter can be achieved through air blow-down with application of a low differential pressure (air or nitrogen) of 5 psi to the filter. Users can apply a buffer chase, but product dilution must be accounted for. At the end of product recovery, sterilizing-grade filters are integrity tested with particle-free, water-based, alcohol (70/30 IPA/ water), or product-based integrity test specifications. For particle-free–water- based or alcohol integrity-test specifications, a filter must be flushed adequately with the test liquid to remove residual product before testing. Product-based integrity-test specifications can be developed through support from filter vendors.

design considerations

Use of Millipak and Millidisk barrier filters in a filtration line is simple and

straightforward (Figures 4 and 5). Similar to all critical applications, important process steps and conditions should be reviewed during system and process design to ensure successful implementation of this application. Verification testing should be performed before implementation of an assembly with barrier filters for product filtration.

Sterilization of Filtration System:

Sterilization renders a system or equipment free of microorganisms and is a critical step, especially for aseptic manufacturing processes. Filters can be sterilized through SIP for Millidisk format (cartridge filters) and autoclaving or gamma irradiation for Millipak (disposable capsule) filters (Figure 6). Thermocouples, radiation dosimeters, and biological indicators serve as the worst-case positions within a filtration system and assembly for validation of sterilization.

Flushing and Wetting Product

Filters: A filtration system may be flushed and wetted to remove extractables after sterilization as well as for product filter integrity testing. The

flushing or wetting liquid passes though the barrier filters to a drain. Flushing/wetting conditions are derived from vendor recommendations (5, 6) and can vary among product filters. Inlet pressures on barrier filters during flushing/wetting procedures should not exceed 0.7 bar. If enhanced wetting of a product filter is required, higher static- hold pressure can be implemented across it. However, the downstream section of that product filter (including the barrier filter) should be isolated during such a high-pressure hold step.

Key Verification Point — Efficacy of Product-Filter Wetting: The efficiency

of wetting a product filter(s) through barrier filters can be verified by performing product filter integrity testing. Results can be compared with filter specifications and past trending.

Key Verification Point — Gas Flow Rate of Barrier Filters After Flushing/ Wetting Procedure: Ensuring that the

hydrophobic membrane in a barrier filter remains dry is critical. Such dryness can be verified in the filter following a flushing/wetting procedure

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during the qualification phase with a low-pressure bubble-stream test. This includes disconnecting barrier filters from their assembly and determining

their gas flow-rate level at 100 mbar (1.5 psi) pressure, then comparing that to the nominal gas flow-rate level of a new filter unit wetted optimally at low

Figure 4: Product filter preparation steps for steamable line, from sterilizing to integrity testing

Product lter preparation steps for steamable line: Testing 1 Sterilizing 2 Wetting 3 Integrity test
Product lter
preparation
steps for
steamable line:
Testing
1 Sterilizing
2
Wetting
3
Integrity test
of product
Close valve
lter
once wetted.
from sterilizing
to integrity
Product
testing
Steam
lter
in place
Atmospheric
Water
(max.
Barrier
Valves
0.5 bar)
lter
Open
Closed
Vent steam through
barrier lter.
Drain water through
barrier lter.
Vent test gas
through barrier lter.
Barrier Filter 4 Drying 5 6 Processing Testing Compressed air Close valve Product lter preparation
Barrier Filter
4 Drying
5
6 Processing
Testing
Compressed air
Close
valve
Product lter
preparation
steps for
steamable line:
once
wetted.
from drying
to processing
70%/30%
IPA/water
Product
Barrier lter
tested o -line
Valves
Open
Closed
Vent air through
barrier lter.

Figure 5: Product filter preparation steps for single-use assemblies, from sterilizing to integrity testing and from drying to process

Product lter preparation steps for single-use assemblies: 1 Sterilizing 2 Wetting 3 Testing Water (max.
Product lter
preparation
steps for
single-use
assemblies:
1 Sterilizing
2 Wetting
3 Testing
Water
(max. 0.5 bar)
Close valve
once wetted.
Air lter
from sterilizing
Product
to integrity
Product
Product
lter
lter
lter
testing
Valves
Barrier
lter
Open
Closed
Sterilized by autoclave
or gamma irradiated as
part of an assembly
Drain water through
barrier lter.
Vent test gas
through barrier lter.
Barrier Filter 4 Drying 5 6 Processing Testing Product Close valve once wetted. Product lter
Barrier Filter
4
Drying
5
6
Processing
Testing
Product
Close valve
once wetted.
Product lter
preparation
steps for
single-use
assemblies:
70%/30%
IPA/Water
from drying
to processing
Barrier lter
tested o -line
Valves
Open
Vent air through
barrier lter.
Closed
Compressed air

32 BioProcess International

14(5)

May 2016

pressure for five minutes. Considering the average hydrophobicity level of polyvinylidene fluoride (PVDF) discs, flow rates in the 30–100% range of nominal rate are characteristic of a “breathing” unit.

Key Verification Point — Using Wetting Medium Apart from Water for

Injection (WFI): Compatibility and intrusion pressure/wettability for the hydrophobic filters within barrier filters should be verified before the filters are used. Wetting hydrophobic filters can reduce their air-flow capacity, leading to increased pressure drop across the filter assembly during integrity testing or product-filter blow-down. Millipak and Millidisk barrier filters validation guides provide

chemical compatibility information summaries (7–9).

integrity testing oF Product Filter

Filtration assembly designers should include strategies to minimize product hold by reducing piping or tubing length and to ensure maximum product recovery. The strategy for integrity testing product filters also should be well thought-out, especially for redundant filtration assemblies. In 2012, Felo and coworkers at MilliporeSigma provided an in-depth look at how product filters can be

integrity tested in a single-use assembly (10). Their strategy can be applied to stainless steel systems as well.

Interference on Product Integrity Testing from Barrier Filters: Barrier

filters are placed downstream of a product filter. To verify the absence of interference, the integrity test result of the product filter both with and without the barrier filters can be compared. Those results should fall within 70 mbar for a bubble-point test

and 5% for diffusion flow.

Failure Mode Test: To simulate a

worst-case scenario (failure-mode test), users can examine how a fully wet barrier filter gas flow rate compromises a product-filter integrity test. Millipak and Millidisk barrier filters can be fully wetted by flushing with WFI at 3 bar.

Adaptation of Troubleshooting

Decision Tree: If a product filter fails its integrity test, users can apply a

troubleshooting decision tree such as the example given in PDA’s Technical Report #26 (3).

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 down a filtration system ranges from 30 minutes to three hours.

Duration of Drying: Exact drying

times should be verified on site and determined during qualification by weight and visual checks. The same time taken to reach the “dry weight” of the assembly will be required for drying the assembly during operation.

Acceptable Applied Pressures:

Typical pressures applied for drying filtration assemblies are 0.5 bar higher than the bubble-point pressure of a product filter. Such pressures should not exceed the maximum allowable pressure of the “weakest link” in an assembly. That might be silicone tubing, a connector, or a barrier filter (4.1 bar for Millipak and Millidisk formats), for example. If the required pressure is greater than what the weakest link allows, then blow-down pressure should be reduced, and an extended drying time can be applied.

Absence of Air-Flow Interference:

Restriction of air flow through fittings, connectors, tubing, or piping used in an assembly should be minimized. As a product filter dries, the air-flow rate will increase and pressure drop across the product filter will decrease.

integrity testing oF Barrier Filters

Barrier filters are integrity tested offline with 70/30 IPA/water as a wetting medium and bubble-point test specification of ≥1,280 mbar (18.5 psi). These filters can be wetted by dynamic flushing or static-soak methods. The wetting procedure of barrier filters can be found in a technical guide (5). A 15-minute static soak can be applied to either Millipak or Millidisk barrier filters. For critical product applications in which resources are readily available, a

Figure 6: Sterilization considerations for Millidisk and Millipak barrier filters

Millidisk Barrier Format Millipak Barrier Format Steam-in-Place Autoclave Sterilization process validation required
Millidisk Barrier Format
Millipak Barrier Format
Steam-in-Place
Autoclave
Sterilization process validation required
Maximum 135 °C for 60 minutes,
up to four times
Steamed in forward direction with 5 psi
(340 mbar) maximum pressure drop
Maximum 123 °C for 90 minutes,
up to three times
Has no vent (self-venting) and serves as
the sterile vent for air and steam of the
autoclaved assembly
Housing should a include housing drain
for condensate removal.
Important Sterilization
Considerations
Filter Format

barrier filter should be integrity tested after the product filter has passed integrity but before product filtration. This minimizes the risk of reprocessing product because of a poor installation or nonintegral barrier filter caused by mishandling. For situations in which resources are limited and product can be reprocessed, barrier filters can be integrity tested after product filtration.

For Best Practices

Barrier filters help enable best practices of aseptic filtration lines for flushing/wetting and preuse integrity testing of product filters. In particular, implementation of Millipak and Millidisk barrier filters is easy and provides flexibility and versatility to the filtration line.

reFerences

1 Annex 1: Manufacture of Sterile

Medicinal Products. Volume 4, EU Guidelines to Good Manufacturing Practice Medicinal Products for Human and Veterinary Use. European Commission: Brussels, Belgium, November 2008; http://ec.europa.eu/health/files/eudralex/

vol-4/2008_11_25_gmp-an1_en.pdf.

2 CBER/CDER/ORA. Sterile Drug Products

Produced By Aseptic Processing: Current Good Manufacturing Practice. US Food and Drug Administration: Rockville, MD, September 2004.

3 Technical Report No. 26: Sterilizing

Filtration of Liquids. Parenteral Drug Association: Bethesda, MD, 2008.

5 P35515 Rev G: Wetting Instructions for

Filter Units with Durapore Membrane. EMD Millipore: Billerica, MA, April 2012.

6 RF1510EN00: Hydrophilic Durapore

Cartridges and Capsules User Guide. EMD Millipore: Billerica, MA, January 2002.

7 VG026 rev 2: Millidisk Cartridge Filter

Units with Hydrophilic Durapore Membrane

Validation Guide. EMD Millipore: December

1999.

8 VG033 Rev D: Millipak Disposable Filter Units Validation Guide. EMD Millipore:

Billerica, MA, May 2012.

9 VG2000EN00: Millidisk Barrier Filter

Validation Guide. EMD Millipore: Billerica, MA, May 2002.

10 Felo M, Oulundsen G, Patil R. Single-

Use Redundant Filtration. BioPharm Int. 25(4) 2012: 38–41.

Corresponding author Yanglin Mok, BE, is a senior process engineer and technical manager of the Biomanufacturing Sciences Network, 1 Science Park Road, #02-10/11 The Capricorn, Singapore 117528; 65-6403-5313, fax 65-6403-5322; yanglin.mok@

merckgroup.com. Lise Besnard, MSc, is a process development scientist at Sanofi Pasteur, 1541 Avenue Marcel Mérieux, 69280 Marcy l’Etoile, France. Terri Love, BSc, is a biomanufacturing engineer; Guillaume Lesage, MSc, is a biosafety technical

consultant; and Priyabrata Pattnaik, PhD, is director of the worldwide vaccine initiative; all with the life-science business of Merck KGaA, Darmstadt, Germany, which operates

as MilliporeSigma in the United States and Canada. Millipak, Millidisk, and Durapore are registered trademarks of MilliporeSigma.

4 ASTM Standard F838-83: Standard

Test Method for Determining Bacterial Retention of Membrane Filters Utilized for Liquid Filtration. American Society for Testing and Materials: Philadelphia, PA, 1983.

For reprints, contact Rhonda Brown of Foster Printing Service, rhondab@fosterprinting.com, 1-866-879-9144 x194.

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