IE20070543U1 - An airlift bioreactor - Google Patents
An airlift bioreactorInfo
- Publication number
- IE20070543U1 IE20070543U1 IE2007/0543A IE20070543A IE20070543U1 IE 20070543 U1 IE20070543 U1 IE 20070543U1 IE 2007/0543 A IE2007/0543 A IE 2007/0543A IE 20070543 A IE20070543 A IE 20070543A IE 20070543 U1 IE20070543 U1 IE 20070543U1
- Authority
- IE
- Ireland
- Prior art keywords
- circulation
- container
- liquid
- tube
- bioreactor
- Prior art date
Links
- 239000007788 liquid Substances 0.000 claims description 36
- 238000005273 aeration Methods 0.000 claims description 10
- 239000012530 fluid Substances 0.000 abstract description 8
- 239000007789 gas Substances 0.000 description 22
- 239000004033 plastic Substances 0.000 description 9
- 229920003023 plastic Polymers 0.000 description 9
- 210000004027 cells Anatomy 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 229920003013 deoxyribonucleic acid Polymers 0.000 description 4
- 239000001963 growth media Substances 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 239000002985 plastic film Substances 0.000 description 4
- 238000004891 communication Methods 0.000 description 3
- 229910001220 stainless steel Inorganic materials 0.000 description 3
- 239000010935 stainless steel Substances 0.000 description 3
- 238000004659 sterilization and disinfection Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 238000010364 biochemical engineering Methods 0.000 description 2
- 230000009089 cytolysis Effects 0.000 description 2
- 238000009630 liquid culture Methods 0.000 description 2
- 230000002934 lysing Effects 0.000 description 2
- 239000002609 media Substances 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- MYMOFIZGZYHOMD-UHFFFAOYSA-N oxygen Chemical compound O=O MYMOFIZGZYHOMD-UHFFFAOYSA-N 0.000 description 2
- 238000005192 partition Methods 0.000 description 2
- 230000001603 reducing Effects 0.000 description 2
- 238000006722 reduction reaction Methods 0.000 description 2
- 230000000630 rising Effects 0.000 description 2
- 241000244206 Nematoda Species 0.000 description 1
- 239000003570 air Substances 0.000 description 1
- 230000001580 bacterial Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- CURLTUGMZLYLDI-UHFFFAOYSA-N carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 230000002759 chromosomal Effects 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 210000003527 eukaryotic cell Anatomy 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 229920002457 flexible plastic Polymers 0.000 description 1
- 238000005755 formation reaction Methods 0.000 description 1
- 238000001415 gene therapy Methods 0.000 description 1
- 230000002706 hydrostatic Effects 0.000 description 1
- 230000036512 infertility Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005020 pharmaceutical industry Methods 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 210000001236 prokaryotic cell Anatomy 0.000 description 1
- 239000012429 reaction media Substances 0.000 description 1
- 230000000979 retarding Effects 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 231100000803 sterility Toxicity 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
Classifications
-
- B01F3/0451—
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/14—Bags
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/06—Nozzles; Sprayers; Spargers; Diffusers
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/06—Nozzles; Sprayers; Spargers; Diffusers
- C12M29/08—Air lift
Abstract
ABSTRACT The present invention is concerned with a fluid circulation apparatus for use as a bioreactor of the like, the apparatus comprising a flexible bag like container within which is disposed at collapsible and at least partially inflatable draft tube which is tethered to a base of the container such that when inflated the draft tube rises off the base and maintains the correct shape as a result of the inflation thereof, and which includes perforations in order to allow the inflation gas to escape and rise upwardly through the draft tube to generate counter current circulation within the container.
Description
Field of the Invention
The present invention provides a liquid circulation apparatus for continuously
mixing a liquid, and a method of mixing such a liquid. More particularly, the
invention provides an apparatus for use as an airlift bioreactor for culturing
prokaryotic and eukaryotic organisms or cells or the like. The bioreactor can for
example be used for growing eukaryotic organisms such as, but not limited to,
nematodes and other worms. In a preferred embodiment, the invention provides a
disposable airlift bioreactor.
Background of the Invention
The IUPAC Compendium of Chemical Terminology defines an airlift bioreactor
as “a bioreactor in which the reaction medium is kept mixed and gassed by
introduction of air or another gas (mixture) at the base ofa column-like reactor
equipped either with a draught tube or another device (e.g. external tube) by which
the reactor volume is separated into a gassed and un—gassed region thus generating
a vertically circulating flow.” The definition refers to internal and external loop
airlift bioreactors, although the present invention is concerned only with the
internal-loop airlift bioreactor.
The majority of airlift bioreactors are traditionally made out of stainless steel and
designed with pressure vessel criteria in mind to sustain the required steam
pressure during sterilisation. The draft tube is also made in stainless steel, is
cylindrically shaped and supported in a vertical position by arms connected to the
inside of the bioreactor vessel. The air inlet is generally positioned just below the
draft tube, so that the gas-liquid mixture rises inside the draft tube, although the
opposite circulation mode is also possible when the air inlet is positioned outside
L
the draught tube. Hence, the overall simplicity of the bioreactor and its
geometrical relationship with the draft tube makes it possible to scale-up
laboratory sized bioreactors without major difficulties to sizes of up to 10,000
litres.
A disposable airlift bioreactor suited to replace the traditional airlift bioreactor
would be especially attractive since it could provide considerable reduction in
capital investment as well as operating costs. The reason for this is the
replacement of the stainless steel bioreactor vessel with a pre-sterilised plastic
bioreactor container or bag which would only be used once; making the otherwise
required sterilisation (e.g. steam) and cleaning process obsolete.
So far, several attempts have been made on disposable airlift bioreactors:
o P. Whitney’s air lift fermenter as part of Patent GB2202549 consists of a
flexible daft tube connected at the top and bottom of the plastic bag
whereby the air inlet is positioned below the daft tube. The problem with
this approach is that suction forces within the draft tube cause it to
collapse, preventing entry ofsubsequent air bubbles.
The invention ‘Air lift femientor formed from flexible plastic sheets‘ as
described in patent EP0343885 uses a ‘divider panel formed of plastic
film’ to divide the plastic bag into two regions and thus creating circulation
in the bag with an air inlet device in one of the regions. In this approach
there is no risk of shear forces inside the draft region causing the dividing
panel to collapse since when the bag is filed, the panel is under tension and
taut. To achieve this configuration, the two plastic films forming the bag
need to be bigger in size than the divider film. This precondition presents
difficulties during fabrication of the bag and requires additional folding to
reduce the overall volume for sterilisation and shipping.
A disposable air lift bioreactor is describes as part of patent US6432698
whereby again a partition is used to create a draft tube. Here the partition
lEa7a5
can either be a divider inserted in the bag or can be created by fusing a
short section of the two sides of the plastic bag together. This approach is
suitable for bags of small volumes, but might not withstand hydrostatic
pressures if applied to larger volumes.
Patent WO2005l 18771 titled ‘Disposable bioreactor systems and methods’
mentions the possibility of a divider inside the bag, more specifically ‘a
sheet of plastic bisecting the chamber of the bag vertically’ to enable
circulation with a gas inlet element. This concept however was already
described in earlier patents (see above) such as EP0343885 and
US6432698
In summary, the two approaches until now for a disposable airlift bioreactor have
been based either on a flexible draft tube or on a divider panel. So far, both of
them have not been suitable to replace the existing traditional airlifi bioreactor.
The draft tube in particular has represented a problem in translating the traditional
airlift bioreactor to a disposable version since bags made out of plastic sheets were
already used to store and transport liquids. Hence, the challenging aspect for a
disposable airlift bioreactor being able to replace traditional bioreactors consists of
a draft tube or other device to provide vertical circulation in the bag.
Nowadays disposable (plastic) bags are increasingly used to store and transfer
solutions required for bioprocessing. The bags can be supplied pre-sterilised and
their shapes and configurations vary extensively. The majority of theses bags are
generally referred to as either two or three dimensional (2D & 3D), whereby a 2D
bag consists of 2 sheets of plastic sealed at their perimeter while a 3D bag consists
mainly of 4 sheets of plastic sealed and cut in a particular form to create a bag with
sides. There are of course other bags available.
It is therefore an object ofthe present invention to provide a liquid circulation
apparatus, and in particular a disposable airlift bioreactor which overcomes the
above mentioned problems of the prior art.
Summary of the invention
According to a first aspect of the present invention there is provided a liquid
circulation apparatus comprising a container; and a circulation guide disposed
internally of the container such as to define a liquid circulation path within the
container; wherein the circulation guide is at least partially inflatable.
Preferably, the circulation guide comprises a draught tube.
Preferably, the circulation guide is flexible.
Preferably, the inflatable part of the circulation guide is of double walled
construction, defining an inflatable cavity therebetween.
Preferably, the container comprises a flexible bag.
Preferably, the circulation guide is tethered to a base of the container.
Preferably, the circulation guide is tethered to the container in a manner which
allows the guide, when inflated or partially inflated, to rise into a substantially
vertical position.
Preferably, the apparatus comprises at least one inflation tube extending between
an exterior of the container and the circulation guide.
Preferably, the circulation guide is wholly inflatable.
Preferably, the apparatus comprises means to aerate, in use, a liquid within the
container in order to generate circulation of the liquid.
Preferably, the aeration means comprises perforations in the circulation guide
which permit, in use, gas to escape in order to generate circulation within the
container.
Preferably, the plurality of perforations are located on an interior portion of the
circulation guide, whereby the sparging of gas into the circulation guide generates
circulation within the container.
Preferably, the aeration means Comprises perforations in the inflation tube.
Preferably, the container comprises at least one inlet and at least one outlet.
Preferably, the container is adapted to receive monitoring devices thereabout.
Preferably, the perforations in the circulation guide and/or the inflation tube are
positioned to cause fluid circulation upwardly about the circulation guide.
Preferably, the apparatus comprises a rigid housing adapted to support the
container therein.
Preferably, the apparatus comprises at least one sampling port.
According to a second aspect of the present invention there is provided a method
of circulating a liquid within a container having a circulation guide therein, the
method comprising aerating the liquid to generate circulation of the liquid about
the circulation guide; and inflating at least a part of the circulation guide in order
to maintain a desired shape and/or orientation of the circulation guide.
Preferably, the method comprises, in the step of aerating the liquid, allowing the
inflation gas to escape from the circulation guide in order to simultaneously inflate
at least part of the circulation guide and aerate the liquid.
lEo7a54s
Preferably, the method comprises, in the step of inflating at least a part of the
Circulation guide, inflating the entire circulation guide.
As used herein, the term “aerate” is intended to mean to supply or charge a liquid
with a gas, especially to charge with air, oxygen, carbon dioxide or the like.
Brief description of the drawings
The present invention will now be described with reference to the accompanying
drawings, in which;
Figure 1 illustrates a perspective schematic illustration of an airlift bioreactor
according to a preferred embodiment of a liquid circulation apparatus of the
present invention;
Figure 2 illustrates the bioreactor of Figure 1 schematically illustrating the
preferred circulation of a culture medium fluid therein;
Figure 3 illustrates a sectioned side elevation of a draught tube forming part of the
bioreactor of Figures 1 and 2;
Figure 4 illustrates a plan view of the draft tube illustrated in Figure 3;
Figure 5 illustrates a sectioned side elevation of an alternative draught tube for use
with the bioreactor of Figures 1 and 2; and
Figure 6 illustrates a plan view of the draught tube illustrated in Figure 5.
M705Detailed description of the drawings
Referring now to Figures 1 and 2 of the accompanying drawings, there is
illustrated a liquid circulation apparatus for particular application as an airlift
bioreactor, generally indicated as 10, which is intended to be used in a disposable
capacity, and in particular for culturing prokaryotic and eukaryotic cells or the
like, in a conventional liquid based cultural medium M (11ot shown in Fig. 1). The
bioreactor 10 comprises a container 12, which is preferably flexible and/or
collapsible, such as a bag made out of plastic sheeting or the like, although in use
it is likely that the container 12 will be supported within a rigid outer casing or
shell or the like (not shown). The container 12 may however be formed of any
other suitable material, and need not be of the shape/configuration shown. In the
preferred embodiment illustrated the container 12 is formed from a sterile flexible
bag where the sterility of the container may be maintained by using current
techniques, e.g. filters, tube sealers & welders, asceptic connectors, etc.
Located internally of the container 12 is a liquid circulation guide in the form of a
draught tube 14 which is open at either end. During normal operation of the
bioreactor 10, the draught tube 14 is located in a substantially vertical orientation
as illustrated, and is suspended off a base 24 of the container 12, such as to define
a circulation path, most preferably a counter current vertical circulation path, for
the liquid M within the container 12, as will be described in detail hereinafter.
The container 12 therefore includes one or more liquid inlet 16, preferably located
at a top of the container 12, and a liquid outlet 18, preferably located at the base
24, although the location and configuration of these may be varied as required.
The container 12 further comprises one or more gas exhausts 20, again preferably
at a top of the container 12, in order to facilitate the venting of aeration gas
introduced into the container 12 during use, again as will be described in detail
hereinafter. The container 12 may also include one or more sensor receptors 22
which enable conventional monitoring devices (not shown) or similar
lEo7o5instrumentation to be used to monitor various operating parameters of the
bioreactor 10, for example the pH of the liquid culture medium M, the oxygen
content, temperature, etc.
The draft tube 14 is tethered to the container 12, and in particular the base 24
thereof, by means of a support 26, although it should be understood that the tube
14 may be secured within the container 12 in any other suitable fashion. The tube
14 is further secured to the base 24 by means of an inflation tube 28 which passes
from an exterior of the container 12, through the base 24, and into fluid
communication with the draught tube 14. In particular, the inflation tube 28
connects with a lower annular portion 30 of the tube 14, which is inflatable,
preferably by means of a double wall arrangement defining an inflatable cavity
therebetween. It should however be appreciated, in particular from the following
description of the operation of the bioreactor 10, that the entire draught tube 14
could be inflatable in similar fashion to the annular portion 30.
In the preferred embodiment illustrated, the bioreactor 10 is provided with aeration
means in the form of perforations or sparge holes 32 located on an interior wall of
the annular portion 30, in order to allow the escape of gas from the annular portion
, to rise upwardly through the tube l4. This aeration, in use, effects circulation
of the fluid M within the container 12, in particular by counter current vertical
circulation upwardly and around the draft tube 14.
Thus, in use, the container 12 of the bioreactor 10 is filled with the liquid culture
medium M, and provided with a quantity of cells or organisms to be cultured.
The inflation tube 28 is then connected to a supply of gas, preferably air, and this
gas is then pumped into at least the annular portion 30 of the draft tube 14. This
results in inflation of the annular portion 30, which therefore results in the opening
or unfurling of the annular portion 30 to its full cross sectional shape. This then
opens up the remainder of the draught tube 14, and also results in the draught tube
rising into a vertical position as illustrated, due to the buoyancy of the air or
lE3?u5é3
other gas inflating same. The air or other gas pumped into the annular portion 30
will then begin to escape through the sparge holes 32, creating a flow of gas
bubbles rising upwardly through the draught tube 14, as illustrated in Figure 2.
This upward aeration through the draught tube 14 sets up a counter-current
circulation within the container 14, as indicated by the directional arrows on
Figure 2. The upward flow of liquid through the draught tube 14 creates suction
forces within the draught tube 14, which would normally act to force the draught
tube 14 to at least partially collapse in on itself, retarding or terminating the
circulation through same. However, by effecting inflation of at least the annular
portion 30, the inflated draught tube 14 provides stability against these suction
forces, thus preventing the draught tube 14 from collapsing. The tethering of the
tube 14 to the base 24 via the support 16, and to a lesser extent by the inflation
tube 28, ensures that the draught tube 14 remains in the position illustrated,
floating off the base 24 to facilitate the counter current circulation therethrough.
By using an inflatable draft tube 14, the draft tube 14 can be flexible, and therefore
collapsible, allowing the bioreactor, when empty, to be collapsed for storage
and/or transport or the like. In addition, when the bioreactor is filled with the
culture medium M, the flexibility of the tube 14, and the container 12, provides a
number of advantages over a rigid arrangement, which advantages are well
documented with respect to other flexible vessels or bags used to store and transfer
solutions required for bioprocessing, not least of which is the reduced cost of
manufacture of the bioreactor 10.
As an alternative to the location of the sparge holes 32 about the annular portion
, it is possible that the section of inflatable tube 28 between the base 24 and the
annular portion 30 could itself be perforated, in order to allow the escape of gas
therefrom, to flow upwardly through the draught tube 14. It should also be
appreciated that the draught tube 14 need not be circular in cross-section, as
illustrated, and could be of any other suitable shape or configuration.
lE‘3705
The bioreactor 10 can also be used as processing equipment for the manufacture of
plasmid DNA. Plasmid DNA is used in the pharmaceutical industry for gene
therapy products and needs to be prepared and isolated after fermentation of the
bacterial cells. Chemical lysis of the cells results in disintegration of the cells and
precipitation of chromosomal DNA, while the plasmid DNA remains in solution.
The bioreactor 10 could therefore be provided with an additional port (not shown)
next to the liquid outlet 18, but extended to reach into the draft tube 14, via which
the chemical lysis gas/solution can be introduced into the bioreactor 10 in a
continuous manner during mixing.
Referring now to Figures 3 and 4, there is illustrated an alternative embodiment of
a draught tube, generally indicated as 114, for use with the bioreactor 10 of the
present invention. The draught tube 1 14 is tethered to the base 24 of the
container, via a support 126. Again, an inflation tube 128 passes from an exterior
of the container into fluid communication with the draught tube 114, which is
inflatable along the full length thereof, and which is again of double wall
configuration in order to allow the inflation thereof. The draught tube 114
includes an inner circumferential ring 40 which is also inflatable, and on an upper
face of which is provided an array of sparge holes 132. The sparge holes 132
permit the escape of the inflation gas from the draught tube 1 14, in order to effect
the vertical aeration of the draught tube 1 14, thereby generating fluid circulation
upwardly through the draft tube 114.
Referring then to Figures 4 and 5, there is illustrated a further alternative
embodiment of a draught tube, generally indicated as 214, for use with the
bioreactor 10 of the present invention. The draught tube 214 is again tethered to
the base 24 via a support 226. An inflation tube 228 extends from an exterior of
the container into fluid communication with the draught tube 214. However,
unlike the draught tube 14 illustrated in Figures 1 and 2, the draught tube 214 does
not include any perforations therein, or any other means to allow the escape of the
$0705inflation gas therefrom. Thus, in order to effect aerated counter current vertical
circulation through the draught tube 214, a sparge tube 50 is provided, which
opens onto the interior of the base 24, and through which may be supplied gas, in
particular air, in order to effect the upward aeration of the draught tube 214.
In an alternative embodiment (not shown) of the present invention, a plurality of
circulation guides may be provided in a single container. The circulation guides
may be placed side by side, stacked one on top of the other, or positioned in any
other suitable or desired formation. Each circulation guide may be supplied with
gas from an inflation tube. In this manner, the aspect ratio of the container may be
adjusted as desired to achieve the desired level of mixing, while achieving a
desired size and shape of the container.
It will therefore be appreciated that the bioreactor 10, and in particular the flexible
and inflatable/collapsible nature of the draft tube 14; 114; 214, enables the entire
bioreactor 10 to be provided as a flexible/collapsible unit with significant
reductions in cost of manufacture and operation. The operation of the bioreactor
of the invention is further improved and refined by combining the use of a gas
to both inflate the draft tube 14 and to effect aeration of the circulation medium M.
The bioreactor 10 therefore extracts a dual functionality from the gas supplied
thereto.
Claims (5)
- l. A liquid circulation apparatus comprising a container; and a circulation guide disposed internally of the container such as to define a liquid circulation path within the container; wherein the circulation guide is at least partially inflatable.
- 2. A liquid circulation apparatus according to claim 1, wherein the circulation guide comprises a draught tube tethered to a base of the container in a manner which allows the guide, when inflated or partially inflated, to rise into a substantially vertical position.
- 3. A liquid circulation apparatus according to any preceding claim, comprising means to aerate, in use, a liquid within the container in order to generate circulation of the liquid, the aeration means comprising perforations in the circulation guide which permit, in use, gas to escape in order to generate circulation within the container.
- 4. A method of circulating a liquid within a container having a circulation guide therein, the method comprising aerating the liquid to generate circulation of the liquid about the circulation guide; and inflating at least a part of the circulation guide in order to maintain a desired shape and/or orientation of the circulation guide.
- 5. The method of claim 4 comprising, in the step of aerating the liquid, allowing the inflation gas to escape from the circulation guide in order to simultaneously inflate at least part of the circulation guide and aerate the liquid.
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
IEIRELAND01/08/2006S2006/0571 |
Publications (2)
Publication Number | Publication Date |
---|---|
IE20070543U1 true IE20070543U1 (en) | 2008-07-23 |
IES84929Y1 IES84929Y1 (en) | 2008-07-23 |
Family
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