U.S. patent application number 14/677670 was filed with the patent office on 2015-07-30 for stirred tank bioreactor.
The applicant listed for this patent is EMD Millipore Corporation. Invention is credited to James E. Kelly, Joseph William Muldoon, Stephen Proulx, James Vigna.
Application Number | 20150210973 14/677670 |
Document ID | / |
Family ID | 41229479 |
Filed Date | 2015-07-30 |
United States Patent
Application |
20150210973 |
Kind Code |
A1 |
Kelly; James E. ; et
al. |
July 30, 2015 |
Stirred Tank Bioreactor
Abstract
The present invention is a disposable, presterilized bioreactor
formed of molded plastic, having a top and body attached to each
other. One or more ports are formed in the top, side and bottom of
the housing. The one or more ports that are below the liquid/air
interface level may be used as sampling ports, access ports for
probes or drains or supply ports for liquids or gases. The
bioreactor provides a direct retrofit for the existing glass or
steel assembly that utilizes the existing support structures and
controls. The molded design overcomes issues of discontinuity, dead
spots and the like due to its fixed dimensions that are built in by
the molding process. Reproducible probe and other equipment
locations are guaranteed through the use of the molded port
features.
Inventors: |
Kelly; James E.; (Melrose,
MA) ; Muldoon; Joseph William; (West Boylston,
MA) ; Proulx; Stephen; (Boxboro, MA) ; Vigna;
James; (North Andover, MA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
EMD Millipore Corporation |
Billerica |
MA |
US |
|
|
Family ID: |
41229479 |
Appl. No.: |
14/677670 |
Filed: |
April 2, 2015 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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|
12387688 |
May 6, 2009 |
8999702 |
|
|
14677670 |
|
|
|
|
61131640 |
Jun 11, 2008 |
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Current U.S.
Class: |
435/289.1 |
Current CPC
Class: |
C12M 23/38 20130101;
C12M 23/20 20130101; C12M 23/28 20130101; C12M 23/46 20130101; C12M
23/48 20130101; C12M 29/00 20130101; C12M 27/00 20130101 |
International
Class: |
C12M 3/00 20060101
C12M003/00; C12M 1/00 20060101 C12M001/00 |
Claims
1) A bioreactor for culturing, fermenting or processing a biomass
comprising: a presterilized, disposable housing made of a rigid
plastic, said housing having a top and a body releasably sealed to
each other, wherein the top and body are sealed to each other by a
gasket interposed between the top and body and a mechanical means
for releasably holding the top and body together, the body having
an interior space, a first set of one or more access ports formed
in the top of the housing and in fluid communication with the
interior of the housing, a second set of one or more access ports
formed in the body and bottom of the housing and in fluid
communication with the interior of the housing wherein at least one
access port of the second set being a valved port in the bottom of
the housing forming an outlet for liquid in the housing, wherein
the valved port further allows for the entrance of gases into the
housing through the port by the valve having a first position to
allow liquid flow, a second position to allow gas flow and a third
position to allow neither liquid nor gas to flow through the valve
and at least one access port of the second set being formed in the
body of the housing so as to allow for the introduction of liquid,
probes or samplers into the interior of the body, the one or more
access ports having a cap to isolate the interior space of the body
from the environment.
2) A bioreactor for culturing, fermenting or processing a biomass
comprising: a presterilized, disposable housing made of a rigid
plastic, said housing having a top and a body releasably sealed to
each other, the body having an interior space, a first set of one
or more access ports formed in the top of the housing, a second set
of one or more access ports formed in the body and bottom of the
housing and in fluid communication with the interior of the housing
wherein at least one access port of the second set being a valved
port in the bottom of the housing forming an outlet for liquid in
the housing, wherein the valved port further allows for the
entrance of gases into the housing by a valve selected to control
flow between gas and liquid through the port, and at least one
access port of the second set being formed in the body of the
housing and in fluid communication with the interior of the housing
so as to allow for the introduction of liquid, probes or samplers
into the interior of the body, the one or more access ports of the
first and second set having a cap to isolate the interior space of
the body from the environment, the top and body are releasably
sealed to each other by a gasket interposed between the top and
body and a mechanical means for holding the top and body together
and wherein the mechanical means is selected from the group
consisting of clips and clamps.
3) A bioreactor for culturing, fermenting or processing a biomass
comprising: a presterilized, disposable housing made of a rigid
plastic, said housing having a top and a body releasably sealed to
each other, the body having an interior space, a first set of one
or more access ports formed in the top of the housing, a second set
of one or more access ports formed in the body and bottom of the
housing and in fluid communication with the interior of the housing
wherein at least one access port of the second set being a valved
port in the bottom of the housing forming an outlet for liquid in
the housing, and wherein the valved port further allows for the
entrance of gases into the housing by a valve selected from the
group consisting of a 3 position valve and Y-shaped tube with
valves on each arm of the Y to control flow between gas and liquid
through the port and at least one access port of the second set
being formed in the body of the housing and in fluid communication
with the interior of the housing so as to allow for the
introduction of liquid, probes or samplers into the interior of the
body, the one or more access ports of the first and second set
having a cap to isolate the interior space of the body from the
environment, the top and body are releasably sealed to each other
by a gasket interposed between the top and body and a mechanical
means for holding the top and body together and wherein the
mechanical means is selected from the group consisting of clips and
clamps and wherein an outer wall of the bottom portion of the
housing has two or more locking tabs and further comprising a stand
having two or more locking features that interact with the locking
tabs of the housing when the housing is inserted into the stand to
hold the housing in the stand in an upright and secure manner.
4) The bioreactor of claim 1 wherein the valve of the valved port
is formed of a 3 position valve and Y-shaped tube with valves on
each arm of the Y to control flow between gas and liquid through
the port.
5) The bioreactor of claim 2 wherein the valved port further allows
for the entrance of gases into the housing by a valve selected from
the group consisting of a 3 position valve and Y-shaped tube with
valves on each arm of the Y to control flow between gas and liquid
through the port.
6) The bioreactor of claim 1 wherein an outer wall of the bottom
portion of the housing has two or more locking tabs and further
comprising a stand having two or more locking features that
interact with the locking tabs of the housing to hold the housing
in the stand in an upright and secure manner.
7) The bioreactor of claim 2 wherein an outer wall of the bottom
portion of the housing has two or more locking tabs and further
comprising a stand having two or more locking features that
interact with the locking tabs of the housing when the housing is
inserted into the stand to hold the housing in the stand in an
upright and secure manner.
Description
CROSS-REFERENCED TO RELATED APPLICATIONS
[0001] This application is a divisional application of U.S.
application Ser. No. 12/387,688, filed May 6, 2009 which claims the
benefit of U.S. Provisional Patent Application No. 61/131,640,
filed on Jun. 11, 2008, the entire contents of which are
incorporated by reference herein.
[0002] The present invention relates to a disposable stirred tank
bioreactor. More particularly, it relates to a stirred tank reactor
formed of a molded plastic having one or more ports molded
therein.
BACKGROUND OF THE INVENTION
[0003] Many small scale bioreactors of the size from about 1 liter
to about 200 liters are formed of glass or steel, preferably
stainless steel. Typical volumes for bench top versions are 2-10
liters. All have a solid body and a removable top sealed to the
body by an o-ring. The top contains ports for probes, sampling, air
sparging, media exchange, and a stir rod for circulation. They are
typically used for culturing or fermenting various organisms such
as plants, bacteria (e.g. E. coli), animal cells (e.g. Chinese
Hamster Ovary (CHO) cells), yeast, mold, etc.
[0004] After each use (typically 3-15 days), the reactor and its
components must be disassembled, cleaned, reassembled, reconfigured
and autoclaved before reuse. This is a time consuming, laborious
process requiring the disassembly and moving of many heavy and/or
small and fragile components. Additionally, one generally needs to
validate the cleaning procedure to ensure that it is done correctly
time after time with the same consistent results. At best after all
the work has been completed, one has rendered the reactor and its
components aseptically clean meaning that contamination can still
occur by residual organisms or advantageous ones that enter through
the aseptic assembly.
[0005] Many designs have attempted to overcome these issues by
using disposable liners in the glass or stainless tank.
[0006] U.S. Pat. No. 6,245,555 suggests using a plastic liner that
is inserted within the existing tank to reduce the amount of
cleaning and increase the level of asepticness. However, it too has
many limitations.
[0007] The liner must conform to the inner surface of the tank in
order to prevent any discontinuity in the circulation within the
device or to prevent the formation of dead spots or pockets in
which material may get trapped and fester or create uneven flow
throughout the system. However wrinkles in the liner still occur
and create the above mentioned problems. All ports are top mounted,
limiting the available area for the different components used in
the bioreactor (feed lines, airlines, stirrer shafts, motors and
journals, sampling ports, probe ports and the like). Additionally,
the top plate is releasably sealed to the liner making only an
aseptic connection. Often times the liner system is limited to an
air sparging system for both gas transfer and circulation. The use
of impeller shafts has been avoided due to the concern that the
shaft or impellers may tear the liner during shipping, storage or
assembly. Additionally, all probes and samplers enter through the
lid or top and have a tube that extends down into the liquid to the
desired level. This means that there is often a long dead leg of
material that needs to be flushed or removed before and after
sampling to ensure that an accurate sample has been obtained.
Lastly, lipids and cholesterols are well-known to bind to many of
the plastics used for such liners.
[0008] What is desired is a disposable tank liner that overcomes
the deficiencies with the current state of the art. The present
invention provides such a device.
SUMMARY OF THE INVENTION
[0009] The present invention is a disposable bioreactor formed of
molded plastic so that it can be rigid or at least semi-rigid and
can be held in a stand or be self standing. The bioreactor is
presterilized and has a top and body sealed to each other. One or
more ports are formed in the top and side of the housing.
Preferably at least one port is below the liquid/air level for the
housing. The one or more ports that are below the liquid/air
interface level may be used as sampling ports or access ports for
probes. Using such a port allows one to take samples without the
need of the dip tube of the prior art eliminating the dead leg and
risk of an improper sample or contaminated sample. Additionally,
the probe does not need to be long in order to fit down to the
desired level in the container. It may simply extend sideways into
the liquid at the desired level. Ports below the liquid level are
an ideal location for the addition of disposable, optical sensors
and provide a means for attaching sensing equipment.
[0010] The invention provides a direct retrofit for the existing
glass or steel assembly that utilizes the existing support
structures, probes for measuring different parameters such as
temperature and pH and controls. The molded design overcomes issues
of discontinuity, dead spots and the like due to its fixed
dimensions that are built in by the molding process. Reproducible
probe and other equipment location is also guaranteed through the
use of the molded port features. Moreover, the rigid bowl is that
it can accept the heating blanket that is used on the glass meaning
that there is no need for an external support or heating jacket.
The molded plastic allows for greater flexibility in material
selection to reduce or eliminate lipid or cholesterol binding.
Preferably the system allows for either an air sparging
gas/circulation system and/or air sparging for gas transfer and a
stirrer/impeller for circulation with out fear of damage to the
container. Molded containers are self supportive and do not require
a support housing as does the flexible liner designs. Additionally
with a molded plastic design heating or cooling blankets can be
directly attached to the molded body whereas in a flexible bag the
blanket must be installed either within or outside of the support
housing. Lastly by having one or more ports formed below the
liquid/air level one can have a drain that allows for the simple
and near complete removal of all liquid when desired.
[0011] It is an object of the present invention to provide a
bioreactor for culturing or processing a biomass formed of a
presterilized, disposable housing made of a plastic selected from
the group consisting of semi-rigid and rigid plastic, said housing
having a top and a body integrally sealed to each other, the body
having an interior space, one or more ports formed in the top and
the body respectively of the housing and in fluid communication
with the interior of the housing, the one or more ports having a
cap to isolate the interior space of the body from the
environment.
[0012] It is another object of the present invention to provide a
bioreactor having two or more ports and at least one port is molded
into the body at a level below a liquid/air interface of the
housing.
[0013] It is a further object of the present invention to provide a
bioreactor having the one or more ports are molded into the top and
body.
[0014] It is an additional object of the present invention to
provide a bioreactor further comprising a stirrer shaft with one or
more paddles mounted within the body of the housing.
[0015] It is another object of the present invention to provide a
bioreactor in which the body includes a port adjacent to a portion
of the body farthest from the top and the port farthest from the
top includes an air diffuser selectively retained to the interior
of the body, the diffuser being formed of a frit selected from the
group consisting plastic, ceramic and metal frits and the port
being connected to a gas line on the exterior portion of the
body.
[0016] It is a further object of the present invention to provide a
bioreactor having one or more ports wherein the one or more ports
of the body are connected to Luer fittings.
[0017] It is an object to provide a bioreactor having a stirrer
shaft with one or more paddles mounted within the body of the
housing and a retainment hub located at the bottom of the body of
the bioreactor to retain and center the shaft.
[0018] These as well as other advantages which will become apparent
form the disclosure below.
In the Drawings
[0019] FIG. 1 shows a first embodiment of the present invention in
perspective view.
[0020] FIG. 2 shows a first embodiment of the body of the present
invention in cross-sectional view.
[0021] FIG. 3 shows a first embodiment of the present invention in
cross-sectional view.
[0022] FIGS. 4A-4C show alternative stirring mechanisms of the
present invention in cross-sectional view.
[0023] FIG. 5 shows an alternative embodiment of the body of the
present invention in perspective view.
[0024] FIG. 6 shows one embodiment of the top according to the
present invention in perspective view.
[0025] FIGS. 7A-7C show one type of fitting and its manner of
attachment to one type of port of the present invention in
perspective view.
[0026] FIG. 8 shows an alternative embodiment of the present
invention.
[0027] FIG. 9 shows one embodiment of the top according to the
present invention in perspective view.
[0028] FIG. 10 shows one embodiment of the bottom of the bioreactor
according to the present invention in perspective view.
[0029] FIG. 11 shows one embodiment of a different support that
interfaces with the bottom of the bioreactor shown in FIG. 10
according to the present invention in perspective view.
[0030] FIG. 12 shows the bioreactor of FIG. 10 connected to the
base of FIG. 11 in perspective view.
DETAILED DESCRIPTION OF THE INVENTION
[0031] FIG. 1 shows an embodiment of the present invention. The
bioreactor 2 is held in a stand 4 which is comprised of several
legs 6 (in this embodiment 3 legs although one continuous leg or 2
large legs or more than 3 legs can also be used) and a support rim
8. As shown the legs 6 may have an optional support piece 10 at or
near the bottom to keep the legs 6 from spreading when the
bioreactor 2 is filled and in the stand 4.
[0032] Depending upon the type of circulation system used the stand
4 may also support the drive mechanism 12 (as shown) for the
circulation mechanism, which typically is a stirrer or paddle
assembly 14 as will be described in greater detail later. In this
particular embodiment, the drive mechanism 12 is a motor and is
mounted to the top of the centered above the top 16 of the
bioreactor 2 by several arms 18 (although 3 are shown alternative
numbers may be used). Other features such as mounting blocks (not
shown) and the like may be formed on the top 16 or support rim 8 to
support the drive mechanism 12. As shown the drive mechanism 12 has
a shaft 20 that can be attached to the stirrer as explained later
herein. Other stands can be used in lieu of the design described
above and will work equally well.
[0033] The bioreactor body 22 has an interior space into which the
fluids, cells, probes and other devices of the bioreactor are at
least partially contained. The body 22 is sealably attached to the
top 16. This may be by a mechanical seal such as a rubber gasket
and clips 24 (as shown) or by a clamp, such as a band clamp or
Ladish or TriClover clamp, mated threads on the top 16 and body 22
and the like. Alternatively, they may be sealed by adhesives or
heat sealing of the top 16 to the body 22 or formed together in one
piece such as in a rotomolding apparatus.
[0034] The body 22 has one or more sidewalls 26 that extend
downwardly from the top 16 and terminate in a closed bottom 28,
preferably having a hemispherical shape. As shown, there is one
sidewall 26 of a circular design which is a retrofit for existing
glass and metal bodies. Alternatively, there can be 3, 4, or more
sidewalls if desired (not shown).
[0035] Preferably, the body is made of a single piece of molded
plastic. Alternatively it may be made of two or more pieces of
plastic that are sealed together such as by heat, glue, or gaskets
(not shown).
[0036] In another alternative arrangement shown in FIG. 5, only the
top 16a and bottom 28a are made of molded plastic and the one side
wall 26a in this embodiment is formed of flexible plastic such as a
plastic film. This still allows for the use of one or more ports in
the device below the liquid/air level. Also notice that the port(s)
such as 32 a may terminate in a hose barb or other connective
feature if desired.
[0037] Also formed in the bioreactor 2 of this embodiment are one
or more ports 30 (in this embodiment there are three types 30a-c
(for a total of 5 ports) formed in the top 16 and one or more ports
32 in the body 22 (in this embodiment there are at least two
different types 32 a-b for a total of seven ports overall in this
embodiment). The top 16 and body 22 may have multiple ports of
similar and/or of different styles to provide one with the number
of ports, of the desired type, in the desired locations throughout
the bioreactor 2. These ports 30, 32 or at least a portion of them
are formed as part of the top 16 and/or body 22. They may be formed
with threads that mate to sealable covers such as closed caps,
gasketed caps with a throughbore within the gasket, or various Luer
fittings. Alternatively, one or more of the ports can be made in
the plastic top 16 and/or body 22 by drilling or burning a hole and
then mounting (such as by heat bonding or adhesives) a port in
place through or around the hole. Many different port styles and
sizes can be accommodated in this invention.
[0038] Ports 30a may be used for liquid or gas entrance or exit or
for probes such as pH probes, thermometers or thermocouples or the
like. Ports 30 b may be used for similar purposes. Port 30c is for
the stirrer shaft described in further detail herein.
Alternatively, if the bioreactor is an airlift design and doesn't
use a stirrer rod, the port 30c may be used to house the airline to
the sparger at or near the bottom of the body or for any other
desired purpose. Ports 32a may be used for sampling of the liquid
or for probes such as pH, temperature, dissolved oxygen, lactose
level, etc as are common on such bioreactors. Ports 32a while shown
as being formed on the sidewall 26 may also be formed in the bottom
if desired as shown in FIG. 2. Port 32b is valved port which can be
used to supply gas to the body 22 and/or as a drain or outlet from
the body. It may serve both functions by attaching a 3 position
valve or Y-shaped tube with valves such as pinch valves on each arm
of the Y to control flow (not shown). One suitable system for the
valve of port 32 b is a LYNX.RTM. connector available from
Millipore Corporation of Billerica, Mass. and as shown in
US2005/0016620.
[0039] Preferably, one or more ports 32 of the body are formed in a
location that is below the normal liquid/gas interface level of the
bioreactor.
[0040] If desired, one or more of the ports 32a or b in FIGS. 1
and/or 2 may be used to provide gases to the body's interior. A
plastic frit such as a POREX.RTM. microporous membrane or ceramic
stone or sintered metal filter may be attached to the inside of the
port within the body to provide the sized gas bubbles desired.
Alternatively, a port 30a in the top 16 may be used to hold a tube
that extends down into the body to provide the gas supply. Again it
may use a frit or ceramic stone or sintered metal filter to provide
the desired bubble size.
[0041] FIG. 3 shows a bioreactor 2 with top 16 and body 22 sealed
to each other and the stirring mechanism 14 in place. The stirring
mechanism is formed of a shaft 40 and one or more paddles 42. The
shaft 40 extends through port 30c and is connected to the shaft 20
of the drive mechanism 12 (not shown). Preferably one or more
o-rings in the port 30c allow for movement of the shaft 40 without
compromising the integrity of the seal within the body 22.
[0042] FIG. 4A shows a stirrer 14 and a retainment hub 50 formed in
the bottom of the body 22. The retainment hub 50 locates and holds
a portion of the shaft so that the shaft does not become dislodged
during shipping, storage or use. Also an optional element, vanes 43
is shown in FIG. 4A. These vanes can be molded into the inner
surface of the body 22 and extend outward into the interior or they
may be separately added after molding. The vanes 43 help to direct
flow and ensure that adequate mixing occurs within the bioreactor
2.
[0043] FIGS. 4B and C show an alternative design to the stirrer 14.
In this embodiment, the gas supply line is built into the stirrer
shaft 40. The shaft 40 has a hollow central bore 44 running through
at least a portion of the shaft. At or near the bottom of the shaft
40 is a gas outlet 46 which may contain a frit, ceramic stone or
sintered metal filter 52 to create the desired bubble size. Gas
enters the bore 44 through gas port 48 in port 30a. One or more gas
conduits 54 connect the inner portion of the port 30a with the bore
44. One or more, in this instance 2 gaskets 56 such as o-rings are
mounted above and/or below the gas port 48 to ensure the gas flows
through the port 48 and into the bore 44 through conduit(s) 54 and
does not escape to the atmosphere or compromise the integrity of
the body. In use the gas flows from a supply into port 48 then into
conduit(s) 54 and down the central bore 44 to the gas outlet 46 in
the shaft and then into the inner volume of the body 22.
[0044] FIG. 6 show one embodiment of the top 16 with some of the
various port configurations and styles that are possible with the
present invention. For example port 30a may be a screw thread style
of port into which a gas or liquid line or probe with matching
mating threads is threaded to create a liquid tight seal.
[0045] FIGS. 7A-C show another port/fitting style. The fitting 31
as shown in FIG. 7C has cam lock 33. This cam lock mates with a
mating cam lock holder 35 in the port 30 (FIG. 7B). The fitting 31
is inserted into port 30 by first aligning the cam locks 33 of the
fitting 31 with the cam opening 35 of the port 30. It is then
pushed downward and rotated to ride over the cam ramp 37 and
terminate and seat itself in the cam lock retainer 39 of the port
30.
[0046] FIG. 8 shows an alternative embodiment of the present
invention incorporating a magnetic mixer 101 within the body 14
driven remotely from a magnetic drive 100 located on the outer
surface of the body 14.
[0047] FIG. 9 shows an alternative top 16 of the invention to that
shown in FIG. 6 in that at least one or more portions of the top 16
incorporates tube clips 202 which can hold the various tubes of the
system in an organized and orderly arrangement. Preferably, the
tube clips 202 are formed on an outer edge 200 of the top 16
although if desired some or all of them may also be attached to the
upper surface of the top (not shown). In this illustrated
embodiment, four clips 202 are shown although more or less may be
used if desired. Preferably they are molded as part of the top 16.
Alternatively, they may be separately attached by melt bonding,
solvent bonding or glues, if desired. Likewise, some or all of the
ports, 30b' and 30b'', can be in the form of hose barbs as shown
rather than screw ports 30 or Luer type fittings as shown in FIG.
6, cam locks of FIG. 7 or other types of fittings discussed below
and commonly used by one of ordinary skill in the art. Tubes
attached to a hose barb fitting such as 30b' can have tie wraps,
connectors such as those taught by US2008/0284163A1 or other
devices or can be adhered to the hose barb to keep the tubes in
place even when under any pressure that may occur in the
system.
[0048] FIG. 10 shows an alternative bottom portion 28 of the
bioreactor 2 to that shown in FIGS. 1-3. In this embodiment,
instead of using the stand 6, the bottom outer wall 204 has two or
more, preferably three or four, locking tabs 206 either molded as a
feature of the bottom outer wall 204 or separately formed and
attached to the bottom outer wall 204 such as by heat bonding,
solvent bonding or glue.
[0049] The locking tabs 206 of the bottom outer wall 204 mate to
corresponding locking features 208 of a bottom support 210 shown in
FIG. 11. The bioreactor is placed into the support 210 and the
locking tabs 206 of the bottom outer wall 204 engage with the
locking features 208 of the support 210. This secures the
bioreactor 2 to the support 210 so that the bioreactor 2 is held
upright in a secure manner as shown in FIG. 12. Additionally, if
desired, one or more open slots 212 can be formed in the support
210 so that any tubing or other features that extend out from the
bioreactor 2 when the bioreactor and support are mated can be
accommodated without removal or pinching. As shown the tabs and
features 206,208 are threaded style devices. However they can be
other forms such as cam locks and cam lock holders as used in some
of the port embodiments discussed above. The locking feature allows
the end user to remove the base from bowl and recycle easily since
the base is not in contact with the culture medium and thus does
not need to be sterilized.
[0050] Fittings such as compression fittings and tube weld, hose
barb and pipe threads, when molded directly into the body, reduce
holdup volume and simplify the system. Such components are well
known and the covers, connectors, septums for sampling (also called
piercable needle ports), check or other valves and the like,
whether Luer type or not; Luer Lok.RTM. fittings; and the like are
readily available for mating with these ports from a variety of
companies such as Value Plastics, Inc of Fort Collins, Colo.
[0051] Suitable polymers which can be used to form the top and body
include but are not limited to polycarbonates, polyesters, nylons.
PTFE resins and other fluoropolymers, acrylic and methacrylic
resins and copolymers, polysulphones, polyethersulphones,
polyarylsulphones, polystyrenes, polyetherimides, nylons,
polyesters, polyethylene terephthalates (PET), polyvinyl chlorides,
chlorinated polyvinyl chlorides, ABS and its alloys and blends,
polyolefins, preferably polyethylenes such as linear low density
polyethylene, low density polyethylene, high density polyethylene,
and ultrahigh molecular weight polyethylene and copolymers thereof,
polypropylene and copolymers thereof and metallocene generated
polyolefins.
[0052] Preferred polymers are polyolefins, in particular
polyethylenes and their copolymers; polystyrenes; and
polycarbonates.
[0053] The top and body may be made of the same polymer or
different polymers as desired.
[0054] Likewise, the polymers may be clear or rendered optically
opaque or light impermeable. When using opaque or light impermeable
polymers, it is preferred that their use be limited to the side
walls so that one may use optical scanners or readers on the bottom
portion to detect the various parameters of the liquid within the
bioreactor.
[0055] Most of the bioreactors of the present invention are
injection molded, but they can be rotoformed in the case that a
jacketed body is required or unique featured are added which are
best accomplished via rotomolding techniques.
[0056] An additional advantage to a molded or rigid formed plastic
body is that heating or cooling blankets can be easily attached to
them.
[0057] In practice, the body is designed as desired, preferably
with a substantially flat open rim one circular sidewall extending
downwardly from the rim and terminating in a rounded bottom as
shown in FIG. 1. The body is preferably made of molded plastic such
as polystyrene, preferably with some or all of the desired ports,
including one or more ports formed in the sidewall and more
preferably with one or more ports formed in the sidewall at a level
below the anticipated liquid/air level. The top is separately
molded with some or all of its desired ports being molded in place.
In many molding applications the ports will actually contain flash
or be closed off by a thin plastic layer that needs to be removed
in order to open up the ports. Alternatively, as discussed above,
the ports may be formed after molding by drilling holes at the
appropriate places and adding port fittings in place in a liquid
tight manner.
[0058] Caps such as various Luer fittings including threaded caps,
Luer based septum covers, Luer Lok caps, closed Luer or other
threaded caps, plugs, tubes attached to the ports formed with hose
barb fittings and the like are attached to the one or more
ports.
[0059] If used, the stirrer shaft 40 with impeller(s) 42 is
inserted into the port 30 in the top 16 and the bottom of the shaft
40 is centered and retained within hub 50. The top 16 is then
sealed to the body 22 such as by the clips 24 or by heat sealing or
adhesive.
[0060] Depending upon the type of probe and the sterilization
technique chosen, one or more of the probes may be added and sealed
in place at this time.
[0061] The closed bioreactor is then sterilized, packaged and sent
to the user.
[0062] The bioreactor may be sterilized by many different
techniques. The most common would be by radiation especially gamma
and to a lesser extent, beta radiation. In this embodiment, many
probes are not gamma stable and would need to be aseptically
assembled into the bioreactor at the user's facility.
[0063] Another method is to use gases such as ethylene oxide. In
this type of sterilization, one could use the various ports to
supply y the gas and then remove it from the interior of the
bioreactor. This may be a dedicated port(s) or it may then be used
for another purpose such as a drain or liquid/air movement.
[0064] Alternatively, the device may be sterilized by autoclaving
with steam and preferably super heated steam and pressure. In this
embodiment, a vent (not shown) to remove the steam would be a
useful addition to one of the ports (30 or 32).
[0065] The sterile device is placed within the stand and the
various connections for air, liquid, probes, sampling, etc are
attached to the device at the appropriate ports. The device is
filled with media to a desired level forming a liquid/air interface
somewhere below where the top 16 is attached to the body 22 to
leave a head space of gas as is common in such devices. At least
one port 32 is below the level of the interface.
[0066] The media is then seeded with the organism to be grown, be
it plant, animal cell (CHO or NSO cells for instance) virus, yeast,
mold or bacteria (such as E. coli) and the liquid is circulated and
air/gases and liquids moved into or out of the device in a manner
to effectively grow the culture inside.
[0067] After a suitable time, the cells may be harvested either by
drawing off the liquid, leaving the cells behind, or in the
instance where the cells need to be ruptured to recover the desired
product by either removing the cells and then rupturing them or
rupturing them in the device and then removing the ruptured mass
for further processing. Additionally, with the vast number of
available ports one could use the device to run a perfusion reactor
in which small amounts of cells or expressed product is removed on
a continual basis for further processing while the cells within the
device continue to grow and make the desired product.
[0068] Once the process is completed the device is drained and all
connections removed and the device ports are sealed. It is then
disposed of properly such as by incineration.
* * * * *