28 Feature

Filtration+Separation May 2006

Gas filtration:
sterile micro-
filtration for
bioreactors
ontinuing with the broad theme of gas filtration, we now look at
C sterilising gas filters - a valuable and indispensable part of
modern biotech and pharmaceutical processes. Jignesh Padia
of QSV Biologics, and Paul Huntington from Cheme
Engineering take us through the process options.

Pharmaceuticals, biotechnology, and healthcare the microbes in the bioreactor, which can be to keep the water vapour above its dew
are high growth segments for the filter market carried over as an aerosol. Bioreactor vent point.
due to the broad growth of aseptic and sterile gas filtration requires some attention
processing. The present cartridge filter market is • Process tank venting:
because it is often saturated with water
about US$2 billion per year with a projected vapour, and this can block the vent filter Process tanks are sterilised in place (SIP)
growth rate of 9 percent. with condensation on the filter membrane with their vent filter installed and the vent
This article is focused on the sterile filtration of or in the filter pores. To prevent wetting of filter is used during cool down – after
gases used for the fermentation process and the filter, either a pre-heater or a heated steaming – to pressurise the tank with sterile
fermenter/tank vent applications. filter housing is used to warm the vent gas air, which breaks the vacuum caused by the
Filter applications for bioreactors and
sterile vessels
Typical gas filtration applications that are seen in
this type of service are:
• Sparger/inlet air filtration:
All bioreactor processes require sterile air or
air/gas mixtures as a raw material. The
amount of air or gas mixture required can
vary for microbial and mammalian cell
culture processes due to their difference in
OUR (oxygen uptake rate) or oxygen
demand. Microbial cells such as E.coli have
a high O2 requirement and can require a
process with at least 30% DO (dissolved
oxygen saturation). In order to meet this
requirement it is not uncommon to sparge
air through the bioreactor at 1-2 VVM
(Volume of air per volume of liquid per
minute).
• Fermenter vent gas filtration:
Vent gas filtration is required for the Pharmaceuticals, biotechnology, and healthcare are high growth segments for the filter market due to the broad
sterility of the process and containment of growth of aseptic and sterile processing.
 Feature 29
Filtration+Separation May 2006

conditions. This is particularly true of vent

Biotech/pharmaceuticals vent filter suppliers
filter sizing and the manufacturer will
require further information about the tank
Manufacturer Brand Name MOC to be vented, to determine the proper sizing
Cuno Microfluor II Hydrophobic PTFE for this application.
Pall Emflon Double-layer hydrophobic PTFE
Millipore Durapore Hydrophobic PVDF Filter housings
Aervent Hydrophobic PTFE
The size of the filter housing is determined
Aerex Hydrophobic PTFE
by the nearest selection of size and number
Sartorius Sartofluor GA Hydrophobic PTFE
of filter cartridges required to meet gas
Domnick Hunter High Flow Tetpor H.T. Expanded PTFE filtration requirements. The selection of the
filter housing type is restricted to the inline
condensing steam. During normal processing liquid challenge test with Brevundimonas style – or the T style – which are given the
conditions the vent filter allows the tank to diminuta ATCC 19146. This evaluation is position of the inlet and outlet connections
breathe sterile air, but precautions need to done based on the complete removal of in relation to the filter cartridge. The T
be taken to ensure that during the process greater than 107 colony forming units (cfu) style housing is the most common choice for
two reasons – it is easier to install; and to
steps the filter does not become wetted or per cm2 of effective filtration area.
configure with a cleaning cap and spray ball
blocked by powders (process materials) from
Filter manufacturers will normally perform for cleaning in place. If you are steaming in
process additions to the tank.
an aerosol challenge test demonstrating place, or your operating pressure is above 15
Cartridge filter construction 100% Brevundimonas diminuta removal, as psig, the filter housing should be purchased
well as a viral and bacteriophage aerosol as a registered pressure vessel built to ASME
The most common type of vent filter used is Section VIII Division 1 code.
challenge test. Note that there is no
the code 7 cartridge filter with the top standardised test procedure for these aerosol
spear. The filter membrane is fabricated in a Integrity test methods for
challenges. Filter manufacturers will also hydrophobic filters
circular pleat, which is bonded to the cage perform a sodium chloride aerosol test using
of the filter cartridge at the top and bottom a condensation nuclei counter (CNC) to To ensure that the filter is performing
creating an inner core. determine the particulate removal rating in correctly, an integrity test (IT) is performed
The support cage is made up of the lower gas. pre and post use. Various IT methods are
mounting lock tabs, the perforated plastic used and acceptable results of these tests are
cage that protects the pleated membrane Filter sizing
and the top end cap that has the bayonet The gas inlet and vent
spear. filters are sized to meet the
In general, filter cartridges are used with the highest flow conditions –
high pressure side on the outside of the at the worst case process
filter, because the filter has the greatest conditions – to be
structural strength in this direction of gas effective throughout the
flow; this is referred to as the forward flow operation of the process.
direction of the cartridge. The sizing should take
into account the following
Hydrophobic filter removal ratings conditions, to determine
the highest flow and worst
Hydrophobic filter cartridges used in such
case process operating
applications have a retention rating of 0.2
conditions before sizing
microns sterilising grade. As per ASTM
the filter:
F838-83 and FDA guidelines, a filter with
this retention rating is evaluated based on a Forward or reverse flow
conditions –
• Normal operating
Features of the ideal air filter
maximum flow and
• Ability to retain organisms through the full
pressure conditions;
range of usage; • Maximum flow and
• High-flow rates at low pressure drop; pressure conditions
during SIP;
• Ease of installation;
• Maximum and dirty
• Ability to in situ integrity test; clean pressure drops.
• Can withstand multiple steaming cycles;
The sizing of the filter is
• Compatible MOC (Material Of best left to the filter
Construction); manufacturer who will
determine the best filter
• Low extractable levels;
selection based on all of
• Regulatory compliant and Validatable. the above operating
30 Feature
Mammalian cell culture
Water intrusion test
Bacterial cell culture
based on the bacterial liquid challenge test
(see hydrophobic filter removal ratings).
Bubble point
In this method, a filter is wetted out using an
alcohol/water mixture (used because the filter
membrane is hydrophobic). The filter is exposed
to increasing air pressure on the upstream side of
the membrane, until the pores in the filter
membrane begin to vent, causing a distinct
change in the gas flow through the filter. The
pressure at which this change occurs is know as
the bubble point and is related to the pore size
distribution in the membrane.
Pressure decay/pressure hold
In this method, a filter is wetted out using
an alcohol/water mixture. The filter is
Filtration+Separation May 2006
exposed to a set pressure upstream of the
membrane. The pressure upstream of the
filter is monitored to determine the flow
rate (based on a ‘know-volume’ upstream of
the filter). The diffusional gas flow rate
across the wetted membrane is directly
related to the pore size distribution in the
membrane.
Note that in both Bubble Point and
Pressure Decay/Hold tests, thorough flushing
of the filter with the alcohol/water mixture
is essential to prevent false IT failures.
Water intrusion test (WIT)
This is a different methodology from the
previous two tests, and the upstream volume
of the membrane is filled with water. This
volume is then exposed to air pressure
upstream of the membrane. After an initial
stabilisation phase, a continuous reduction
in upstream water volume can be observed.
This reduction in water volume is caused by
water diffusing through the membrane, and
can be measured by either monitoring the
pressure drop upstream of the filter, or the
gas flow required to maintain pressure
upstream of the filter.
It is essential that the membrane cartridge is
allowed to cool down to ambient conditions
before the water intrusion IT is performed
on the filter and housing. Cold water
contacting the warm membrane can result
in localised water penetration – resulting in
false positive water intrusion result.
Sterilise in place (SIP)
Sterile vent filters need to be sterilised in
place with the tank – to be truly sterile. The
maximum process conditions that most
filters should be subjected to vary from
manufacturer to manufacturer but are
typically 125oC for 30 minutes at 15 psid in
the forward flow direction and 7 psid in the
reverse.
Always confirm the suitability of the filter
for the SIP conditions – with the filter
manufacturer – before use.
Steaming in the reverse flow direction
requires that the number of sterilisations be
reduced from the forward flow duty cycles,
and filters are more sensitive to pressure
variations in reverse flow.
For SIP the correct rated housing, piping,
and valves should be selected that meets the
requirements of ASME BPE 2004, ASME
Process Piping B31.3 and the ASME Boiler
and Pressure Vessel Code Section VIII Div 1.
Drain lines on the housings must have
sanitary steam traps that can handle the
 Feature 31
Filtration+Separation May 2006

condensate load, provide ports for
temperature monitoring points and, install
sections to hold and recover spore strips for
sterilisation studies.
It is critical to review how you are going to
perform a post SIP IT test without breaking
sterility. This is most usually achieved by IT
of the filter, once the main vessel has been
sterilised and can be used as a sterile
reservoir for the filter during IT.
Clean in place (CIP)
Once vent filters housings are larger than
one round, they are difficult to remove
manually and consideration should be given
to cleaning them in place. To obtain the
best possible CIP conditions, use one of the
modern cleanable filter housings that has a
machine base that eliminates the tube sheet,
and a spray ball that can contact all wetted
parts.
Ensure that there is an adequate supply of
CIP fluid at sufficient flow and pressure to
meet the housing spray ball requirement,
and sufficient drainage to handle the CIP
flow without allowing it to collect in the
base of the housing.
the filter works, and is suitable for their
application.
The manufacturer is responsible for
providing information in support of the end
user’s requirements for details on – the
filter’s performance; materials of
construction, integrity data correlations to
microbial retention; data on the filter’s
range of application; and the validation of
the integrity testing of the integrity tester.
The filter manufacturers produce validation
guides that will include the test methods,
test data and support data that you will use
to support your validation.
As part of your validation effort, the key
points that need to be addressed are:
• Validation of the bacterial retentive
properties of the filter on the actual
product;
• Proving that the filter remains integral at
the end of the filtration run;
• Validating the integrity tester used for
integrity testing of the filter;
• Proving that the filter does not release
harmful extractables into the product;
• Prove that the filter does not adsorb or
absorb components of the product
formulation.
Summary
Membrane cartridge filters are a very cost-
effective way to produce sterile gas streams
for bioreactors and sterile process tanks.
They can be effective when applied
correctly, taking into account the
application and its process conditions.
Finally the filter xmust be validated – and
this process involves the physical, chemical
and biological testing of the filter under
extreme process conditions to provide
assurance that the filters (when correctly
used and integrity tested) will provide a
•
consistently sterile gas stream.
Authors:
Jignesh Padia [www.qsvbiologics.com]
– Process development associate
Paul Huntington [Cheme Engineering Inc]

Filter material compatibility

Filter material compatibility with the
process application needs to be tested before
using membrane filters in a Good
Manufacturing Practices (GMP)
environment.
Incompatibilities between the filter and the
process can arise from one or all of the
following – chemical nature of the gas or
aerosols; adsorption on the filter;
temperature; or pressure.
It is the responsibility of the end user to
verify that the process is compatible with
the materials of construction of the
cartridge filter, and that the process does
not cause the degradation or extraction of
the filter material into their product – by
using an extractable testing program. Filters
also need to be non pyrogenic according to
USP and must meet the requirement for
bacterial endotoxins of
<0.25 EU/mL. They should also pass the
USP Plastics Class VI Test for the injection
testing of extracts and implantation of the
materials used to make the filter.

Validation
The user and the filter manufacturer share
the responsibility for the validation of the
filter application. The user is responsible
for final validation testing to ensure that