Patent ID: 12227730

DETAILED DESCRIPTION

Reference will be made below in detail to exemplary embodiments of the invention, examples of which are illustrated in the accompanying drawings. Wherever possible, the same reference characters used throughout the drawings refer to the same or like parts.

Embodiments of the invention relate to pump devices and related methods to add or remove a known volume of fluid to or from a fluidic vessel in a functionally closed manner. In particular, embodiments of the invention center around a graduated chamber, stopcocks, check valves, sterile filters, and sterile-weldable tubing, but is generalizable to other similar or equivalent embodiments. In its simplest form the pump device may be manually operated, but the operation could be automated without fundamentally altering the embodiments of the invention.

The embodiments of the invention disclosed herein, therefore, address the challenge of the prior art by utilizing sterile-weldable tubing to make the connections (though this aspect is generalizable to other means of aseptic connections, such as self-wiping connectors), and then using a syringe outboard of a sterile filter as a pump to draw fluid from a bioreactor or culture vessel, and then push it to the sample collection vessel or receptacle. Alternately, in an embodiment, the graduated chamber could itself be used as the collection vessel. The invention described herein allows for either stopcocks (which must be actuated, either manually or mechatronically), or check valves (which require no intervention) to ensure fluid flow in only one direction.

As disclosed hereinafter, the simplest embodiment includes a graduated chamber into which the fluid is first collected (graduated so that the user may visually gauge and control how much sample is pulled), and from which the fluid is discharged to a collection vessel. It is envisioned that the syringe action could be automated, and the sample volume metered that way (rather than by eyeball) without departing from the broader aspects of the invention.

This concept can be applied both to push fluid into a closed vessel as well as to remove it from said vessel. The only difference in practice would be the direction of the check valves. In one embodiment, in which the closed vessel is a cell culture chamber, a pair of such devices (of appropriate volumetric capacity) could be used to manually, semi-manually, or automatically effect perfusion (i.e., balanced simultaneous and continual addition of fresh media and removal of spent media) into and out of said vessel.

As used herein the phrase, “biological samples” refers to any particle(s), substance(s), extract(s), mixture(s), and/or assembly(ies) derived from or corresponding to one or more organisms, cells, and/or viruses. As will be appreciated, cells which may be cultured in an automated cell management system includes one or more cell types including, but not limited to, animal cells, insect cells, mammalian cells, human cells, transgenic cells, genetically engineered cells, transformed cells, cell lines, plant cells, anchorage-dependent cells, anchorage-independent cells, and other cells capable of being cultured in vitro. The biological sample also includes additional components to facilitate analysis, such as fluid (for example, water), buffer, culture nutrients, salt, other reagents, dyes, and the like. Accordingly, the biological sample may include one or more cells disposed in a growth medium and/or another suitable fluid medium.

As used herein, the term “sterile” or “sterile environment” refers to an environment that is substantially free of unintended microorganisms.

Moreover, as used herein, the term “sample source” refers to any suitable apparatus, such as a large fermentation chamber, bioreactor, bioreactor vessel and/or culture vessel, for growing organisms such as bacteria or yeast under controlled conditions for production of substances such as pharmaceuticals, antibodies, or vaccines, or for the bioconversion of organic waste. Further, the term “sample source” includes vessels for both aerobic and anaerobic cultivation of microbial, animal, insect and plant cells, and thus encompassing a fermentor.

Further, as used herein, “cell culture” entails growth, maintenance, differentiation, transfection, or propagation of cells, tissues, or their products.

Also, as used herein, the term “biological inoculum” refers to cell culture, cells suspended in growth media, suspension cells, cell aggregates, cells attached to beads and suspended in the growth media, and the like. Further, the term “biological inoculum” also refers to various cell types, such as, but not limited to, mammalian cell types (for example, Chinese Hamster Ovary (CHO), human embryonic kidney (HEK), human embryonic stem cells (hESC), primary human cells, T-cells, and the like), insect cell types, plant cell types, microbial cell types, and the like.

Moreover, as used herein, the phrase “growth medium” or “growth media” is used to refer to a liquid solution used to provide nutrients (for example, vitamins, amino acids, essential nutrients, salts, and the like) and properties (for example, similarity, buffering) to maintain living cells (or living cells in a tissue) and support their growth. Commercially available tissue growth medium is known to those skilled in the art. The phrase, “cell growth medium” as used herein means tissue growth medium that has been incubated with cultured cells in forming a cell culture; and more preferably refers to tissue growth medium that further includes substances secreted, excreted or released by cultured cells, or other compositional and/or physical changes that occur in the medium resulting from culturing the cells in the presence of the tissue growth medium.

Additionally, as used herein, the term “sampling instance” may be used to refer to an event of drawing a sample from a sample source at a given instance in time.

Further, as used herein, the term “aseptic sampling” refers to sampling while preventing entry of contamination or external impurities in the sample source or associated components.

Also, as used herein, the term “tubing” may refer to at least a portion of one or more of a sampling conduit, a recovery conduit, and one or more sub-conduits.

As used herein, the term “fluid communication” refers to a relationship between two components by which fluid can be permitted to flow from one component to the other.

FIG.1illustrates a sampling assembly10(also referred to herein as sampling system10) configured for aseptic sampling of one or more samples from a sample source12. In certain embodiments, the sample source12may be a suitable culture vessel that is configured for cell culture, such as, but not limited to, cell expansion and growth. Further, the sample source12may be configured to house a biological inoculum. In some embodiments, aseptic sampling may be performed to monitor the cell culture process occurring in the sample source12. A sampling performed at a given time may be referred to as a sampling instance. In one embodiment, a plurality of sampling instances may be performed using the sampling assembly10in a time efficient and aseptic fashion.

As shown inFIG.1, the sampling assembly10(also referred to herein as sampling system10) includes a graduated sampling chamber14configured for fluid connection to the sample source12, such as via connection to a port16on the sample source12, and a pump device such as a syringe18configured for fluid connection to the sampling chamber14. The size of the sampling chamber14may be selected according to the particular application or sampling operation carried out, and may range from about 0-5 mL, or from about 1-3 mL, although smaller or larger collection volumes are also envisioned by utilizing an appropriately sized chamber. In particular, it is contemplated that to carry out certain processes, the volume of the sampling chamber14may be tens or hundreds of milliliters. The graduated chamber14, by its definition, has a plurality of graduations or markings enabling a user to see the amount of fluid contained within the sampling chamber14. In an embodiment, the sampling assembly10includes a first three way valve20intermediate the graduated sampling chamber14and the sample source12, such that the graduated sampling chamber14can be selectively placed in fluid communication with the sample source12and/or a receptacle22connected to the valve20via tubing24, as discussed in detail hereinafter. In an embodiment, the receptacle22is a vacutainer collection tube, and the tubing24is a length of weldable PVC or similar material. The sampling assembly10additionally includes a second three way valve26intermediate the graduated sampling chamber14and the syringe18, such that the graduated chamber14can be selectively placed in fluid communication with the syringe18and/or with another device through secondary port28of the three way valve26.

WhileFIG.1illustrates the use of a syringe16, other manual, semi-automatic, or automatic pump devices (e.g., a motorized pump) may also be utilized without departing from the broader aspects of the invention. In an embodiment, the valves20,26may be pinch valves or stopcocks, although other types of valves known in the art configured to provide for multiple flowpaths between components may also be utilized. It is contemplated that the valves20,26can be manually controlled or controlled automatically via an actuator.

With further reference toFIG.1, the sampling assembly10also includes a sterile air filter30positioned in the flowpath between the pinch valve26and the syringe18, as well as a sterile air filter32associated with port28(e.g., within a flowpath connecting an auxiliary device (not shown) to the pinch valve26via port28).

In use during a sampling operation, the first valve20is controlled to a position to place the sample source12in fluid communication with the graduated sampling chamber14, while the second valve26is controlled to a position to place the syringe18in fluid communication with the sampling chamber (via port34). The syringe18is then utilized to draw or pull a desired volume of fluid from the sample source12into the graduated sampling chamber14. As indicated above, the graduations on the chamber14are utilized to easily verify when a desired amount of fluid has been drawn into the chamber14. Once a desired volume of fluid is present in the chamber14, the valve26is controlled to place the port28in fluid communication with the chamber14, and the valve20is actuated to fluidly isolate the sample source12from the chamber14, and to place the receptacle22in fluid communication with the chamber14. In the case where the receptacle22is a vacutainer, upon enabling fluid connection between the receptacle22and the chamber14, the vacuum environment within the receptacle22pulls the fluid within the chamber14into the tubing24and receptacle22, displacing it with air let in through the sterile air filter32and port28in the valve26. In such case, it is envisioned that the receptacle22is large enough to completely empty the chamber14plus the tubing24. In an embodiment, the tubing24may be selected to be long enough so as to conveniently position the receptacle22at a location where it can be easily accessed for sampling and analysis.

In addition to verifying by sight using the graduated markings, to determine the amount of fluid drawn into the chamber14, in other embodiments, the fill volume of the chamber14may be ascertained by automated optical sensing methods and/or by weight.

In an embodiment, the receptacle22need not be a vacutainer. In such embodiments, a syringe or other pump device fluidly connected to chamber14through valve26can be utilized to push the volume of fluid present in the chamber14to the receptacle22. In particular, air injected through either port28,34, for example by syringe18, may be utilized to push the volume of fluid all the way to the receptacle22. In either case, the presence of the sterile air filters30,32ensures that any air entering the system10is sterile. Once the sample is collected in the receptacle22, the receptacle22can be sterile-welded off and replaced with another receptacle for further sample collection.

In an embodiment, prior to connecting another sample collection receptacle a purge step may be carried out to clear the chamber14and tubing24, if desired. This purge step may be carried out in a variety of ways. In one embodiment, a waste flush receptacle (not shown) may be connected to the port36on the valve20via tubing24so that the holding chamber14, valve passages and/or tubing24can be flushed before further sample collection. For example, in an embodiment, the sampling assembly10may include a reservoir of sterile fluid such as water or saline connected to port28(or another port, not shown) that is utilized to flush the chamber14and the tubing24once the sample has been collected, flushing with air when done, and then connecting a new sample collection receptacle to the tubing24. A similar purge or flushing process is disclosed below in connection withFIGS.4and5.

In another embodiment, another vacutainer may be connected to tubing24so that air can be drawn into the chamber14through one of the sterile air filters30,32and passed through the chamber14and tubing24. In yet another embodiment, a syringe of saline or other fluid may be connected to one of the ports28,34of valve26and actuated to flood the chamber14with sterile saline. A vacutainer connected to tubing24may then be utilized in a manner similar to that disclosed above to draw the saline from the chamber14, through tubing24and into the vacutainer. In the case where a vacutainer is not utilized, sterile air can be injected through the valve26to push the saline from the chamber14into the purge receptacle via line24. In yet another embodiment, the valves20,26may be controlled so that a syringe connected to valve26is in fluid communication with the chamber14, and so that chamber14is in fluid communication with the sample source12. The syringe can then be utilized to push fluid inboard of the valve20back into the sample source12by gravity. In an embodiment, the vertical orientation of the graduated chamber14assists in measuring and emptying. It is contemplated that in some embodiments, a check valve may be positioned inboard of the valve20for preventing fluid from being inadvertently pushed back into the sample source12.

In connection with the above, in an embodiment, the second valve26may be omitted in favor of a single port. A user can then just unscrew the syringe18after using it to draw the fluid from the sample source12into the chamber14to let air in through the sterile air filter.

Referring now toFIG.2, a bioprocessing assembly100(also referred to herein as bioprocessing system100or sampling assembly100) according to another embodiment of the invention is shown. The bioprocessing assembly100includes a culture vessel110which may be, for example, a static culture vessel containing a population of cells. As illustrated, the vessel110includes two tubing tails112,114(e.g., connected to opposing ends of the vessel110), and two manual syringe pumps116,118welded to the tubing tails112,114, respectively. The manual syringe pumps116,118are generally similar in configuration to those described above in connection withFIG.1. In particular, manual syringe pumps116,118each include a graduated chamber120having a plurality of graduations thereon for visually determining a volume of fluid within the chamber120. Each of the pumps116,118also include a pump device such as a manual syringe122configured for fluid connection to an upper end of the chamber120. A sterile air filer124is disposed intermediate the chamber120and the syringe122. In an embodiment, the sterile air filer124is permanently attached to the chamber120. In other embodiments, the sterile air filter124may be connected via a luer taper to the syringe122.

The graduated chambers120of the first and second syringe pumps116,118are in fluid communication with the vessel110via the tubing tails112,114, respectively. In an embodiment, a check valve126,128is positioned along the fluid pathway (i.e., along the tubing tails112,114) between the vessel110and the chambers of the syringe pumps116,118, respectively, permitting only unidirectional flow of fluid as indicated by the arrows.

As further shown inFIG.2, the bioprocessing assembly100may further include a source reservoir130fluidly connected to the graduated chamber120of the first syringe pump116via tubing132, and a waste reservoir134fluidly connected to the graduated chamber120of the second syringe pump118via tubing136. As discussed hereinafter, the source reservoir130may contain various fluids for use in bioprocessing operations such as, for example, fresh media, coating solution, virus, etc. As illustrated, both tubing runs132,136may be fitted with check valves138,140which permit only unidirectional flow of fluid (e.g., from the fluid source130to the chamber120of the first syringe pump116, and from the chamber120of the second syringe pump118to the waste reservoir134) as indicated by the arrows. In an embodiment, the various components of the assembly100may be fluidly interconnected at sterile weld points142along the tubing lengths.

As will be appreciated, the syringe pump116(and the syringe122thereof) allows for the aseptic transfer of fluid from the source reservoir130to the vessel110and/or from the vessel110to a waste receptacle134or other downstream bag or receptacle. In particular, syringe116may be utilized in a manner similar to that described above in connection withFIG.1to draw a fluid from the source reservoir130, through the tubing132, and into the graduated collection chamber120of the first syringe pump116. As disclosed above, the graduations on the chamber120allow a user to precisely control the amount of fluid drawn into the chamber. The same or different syringe or pump can then be used to push air through the sterile air filter124, thereby pushing the fluid within the chamber120through tubing112and into the vessel110. In an embodiment, the fluid may be moved from the chamber120of the first syringe pump116to the vessel110under force of gravity. Syringe pump118can be operated in similar manner to move fluid from the vessel110to the waste receptacle134, by drawing fluid out of vessel110and into the chamber120through tubing114via actuation of the syringe122, and then by pushing air into the chamber120through sterile air filter124to move the fluid from the chamber120to waste receptacle134through tubing136. In an embodiment, the fluid may be moved from the chamber120of the second syringe pump118to the waste receptacle134under force of gravity.

The assembly100of the invention shown inFIG.2, therefore, can be utilized to carry out a variety of bioprocessing operations, such as perfusion. Where manual syringe pumps are utilized, it is envisioned that such perfusion would be sporadic, pulse perfusion (carried out at selected intervals). For example, using the assembly100, manual pulsed perfusion may be carried out at an approximate rate of 0.5 L/day (which would require, for example, 5 100 mL volumes per day). It is contemplated, however, that perfusion can be carried out automatically, and in a continuous or pulsed manner, using a mechanical pump instead of a manual syringe. In particular, whileFIG.2illustrates the use of a manual syringe, other manual, semi-automatic, or automatic pump devices (e.g., a motorized pump) may also be utilized without departing from the broader aspects of the invention.

As with the embodiment ofFIG.1, the syringes122of the pumps116,118serve solely as a means to create a vacuum to draw fluid into the graduated chambers120of the pumps116,118, respectively, or to create positive pressure to dispel fluid. Accordingly, with this configuration there are no worries of contamination as the syringes122are not part of the fluid paths (i.e., they never come into contact with the fluid moving into or out of the source reservoir130, culture vessel110or waste receptacle134. Accordingly, in any of the embodiments disclosed herein, the syringe pumps and components thereof need not even be sterile, as they never come into contact the bioprocess fluid. In this manner, the assembly100provides a fully closed manual culture system that obviates sterility and contamination issues that have not been solved using existing systems and methods.

In addition to the configuration of the assembly100shown inFIG.2, it is further contemplated that the assembly100may include pump devices similar to pump devices116,118or that disclosed above in connection withFIG.1for aseptically pulling samples or to make small volume additions to the vessel110. In this way, the assembly100can be utilized to draw a sample from the vessel110for testing of the fluid (e.g., for determining cell density).

Turning now toFIG.3, an aseptic sampling assembly200in the context of a recirculating loop according to another embodiment of the invention is illustrated. The sampling assembly200includes a bioreactor vessel210in the form of, for example, a static culture vessel containing a population of cells, and a recirculating loop212(e.g., formed from tubing and having a pump (not shown) to circulate the fluid) having a first end fluidly connected to an outlet of the vessel210for receiving fluid from the vessel210, and a second end also fluidly connected to the vessel210for returning the fluid to the vessel210. The sampling assembly200further includes a manual syringe pump116fluidly connected to the recirculating loop212. In an embodiment the manual syringe pump116is similar or identical to the manual syringe pump116,118ofFIG.2, where like reference numerals designate like parts. A length of tubing216having an optional check valve218serves to connect the chamber120to the recirculating loop212. In an embodiment, the syringe pump116is fluidly connected to the recirculating loop212via a sterile connector214. The sterile connector214may be, for example, a reusable sterile connector. In an embodiment, the reusable half of the sterile connector214may be part of the kit (containing the vessel210and recirculating loop), while a single-use mating portion may be part of a sampling accessory (comprising the manual syringe pump116including chamber120, tubing216and check valve218).

In use, cells reside in the vessel210, but for a sampling event, are recirculated through the loop212to ensure homogeneity. A sample can then be manually pulled from the recirculating fluid using syringe pump116in the manner hereinbefore described. As indicated above, the manual syringe pump116connects aseptically to the loop212via the reusable aseptic or sterile connector214. To collect a sample, a user would first connect the syringe pump116to the recirculating loop212via the connector214. Then, through manual action of the syringe122, a sample is pulled through the tubing216, past the check valve218, and into the graduated chamber120. As disclosed above, the chamber120is graduated so as to allow a user to see the precise volume being pulled, and provides a means to pull a sample of any desired volume. As also indicated above, the syringe122is attached to the chamber120via an intermediate sterile air filter124to mitigate the risk of accidentally back-flushing non-sterile air into the system. This risk can be further mitigated by inclusion of check valve218along tubing line216. Omitting the check valve218, however, allows for aseptic backflushing into the loop212in the manner described above in connection withFIG.1.

WhileFIG.3illustrates the use of a manual syringe, other manual, semi-automatic, or automatic pump devices (e.g., a motorized pump) may also be utilized to carry out the sampling operations disclosed herein, without departing from the broader aspects of the invention. Moreover, while the system200ofFIG.3discloses the use of a reusable connector214, it is contemplated that multiple ports along the recirculation loop212may, alternatively, be employed for the withdrawal of multiple samples.

With reference toFIGS.4and5, a sampling assembly300for use in a bioprocessing system according to an embodiment of the invention is shown. The sampling assembly300is similar to the syringe pump disclosed above in connection withFIGS.1-3, and includes a bioprocessing vessel such as, for example, a single use bioreactor310containing a fluid (e.g., a cell culture). The sampling assembly300includes a graduated chamber312fluidly connected to the single use bioreactor310, and a syringe314fluidly connected to the graduated chamber312. A sterile air filter316is disposed between the syringe314and the graduated chamber312for the purpose hereinbefore described. The sampling assembly300is operable in the manner described above in connection withFIG.1, to draw a sample from the single use bioreactor310into the graduated chamber312.

As illustrated inFIGS.4and5, in an embodiment, the single use bioreactor is fluidly connected to the chamber312at one end of the chamber312(i.e. top or bottom), while the syringe314is fluidly connected to the chamber312at an opposite end of the chamber312(i.e., bottom or top). With particular reference toFIG.4, to pull a sample, the chamber is oriented such that fluid is drawn from the bioreactor vessel310into the bottom of the chamber310(while the syringe314draws air out of the chamber310from the top). This process is carried out until a desired amount of fluid is present in the chamber310, as visually indicated to a user by the markings on the chamber310. With particular reference toFIG.5, to purge or flush the chamber310and flow lines (such as the process contemplated in the discussed ofFIG.1), in an embodiment, the sampling assembly300is inverted such that the syringe and connection to the chamber310is located vertically below the connection point of the bioreactor vessel310to the chamber. The syringe314is then depressed. Air will bubble up through the fluid within the graduated chamber310and back down the flow lines/tubing to the bioreactor vessel310, thereby purging the lines.

Turning now toFIG.6, a sampling assembly400according to another embodiment of the invention is shown. The sampling assembly400is similar to sampling assembly300ofFIGS.4and5, where like reference numerals designate like parts. Rather than having the syringe314and bioreactor vessel310connect to the graduated chamber312at opposite ends thereof, however, the syringe314and bioreactor vessel310are both fluidly connected to an upper end portion of the graduated chamber312(on opposing sides of the chamber312). In such embodiment, the graduated chamber312includes a baffle410or divider that extends downwardly into the interior area of the chamber312from the top thereof.

In use, to pull a sample from the bioreactor vessel310, the syringe314is used in the manner disclosed above to pull fluid into the graduated chamber312. Fluid will fill the chamber312, through port420, entering from the top port connection to the bioreactor vessel310and collecting at the bottom therefore due to the force of gravity. The baffle410serves to prevent pulling fluid straight across to the syringe314. To purge or flush the flow lines, the syringe314is depressed to push air through the sterile air filter316and port430and into the graduated chamber312. The air will circuit around the baffle410, through the fluid (in the case that the fluid extends to the baffle), and back down the line to the bioreactor vessel310.

With reference toFIGS.7and8, in an embodiment, the port connections420,430to the bioreactor vessel310and syringe314, respectively, may be integrated into a cap440of the graduated chamber312, which is configured to be received at the top the graduated chamber312. In this respect, and as shown inFIGS.7and8, the graduated chamber312includes a rubber stopper442fitted in the neck of the chamber312that is designed to frictionally engage the cap440for removable coupling of the cap440to the chamber312. While a friction fit is illustrated inFIGS.7and8, it is contemplated that other means of releasable connection such as threaded engagement, a bayonet mount, and the like may also be utilized without departing from the broader aspects of the invention. As will be appreciated, this configuration allows the cap440to be removed so as to access the collected sample (rather than having to push it to a separate collection receptacle as disclosed inFIG.1.

Referring now toFIGS.9and10, a sampling assembly500for use in a bioprocessing system according to an embodiment of the invention is shown. The sampling assembly500is similar to those described above, and includes a bioprocessing vessel such as, for example, a single use bioreactor510containing a fluid (e.g., a cell culture). The sampling assembly500includes a graduated chamber512fluidly connected to the single use bioreactor510, and a syringe514fluidly connected to the graduated chamber512. A sterile air filter516is disposed between the syringe514and the graduated chamber512for the purpose hereinbefore described. As shown inFIG.9, the bioreactor vessel510is fluidly connected to the graduated chamber512at one end thereof, while the syringe514is fluidly connected to the same end of the graduated chamber512via a first tubing line518and to an opposite end of the graduated chamber via a second tubing line520. As illustrated, the first tubing line518includes a check valve522that only permits flow out of the bioreactor vessel510, while the second tubing line520includes a check valve524that only permits flow into the bioreactor vessel510.

With particular reference toFIG.9, in use, to obtain a sample from the bioreactor vessel510, a user uses the syringe514to pull fluid into sampling chamber512in the manner hereinbefore described. Fluid will fill the chamber from the bottom, as shown inFIG.9. With reference toFIG.10, to flush or purge the lines, the sampling assembly500is inverted similar to the manner described above in connection withFIGS.4and5. The syringe514is then depressed to force air into the chamber512through the sterile air filter516and through the second tubing line520. The check valves522,524will force the air down the external bypass path (i.e., through line520), over the fluid and sweep the line back into the bioreactor vessel510.

It is contemplated that any of the embodiments ofFIGS.4-10may be integrated into the assemblies/systems ofFIGS.1-3. Moreover, similar toFIGS.1-3, while the embodiments ofFIGS.4-10illustrate the use of the use of a syringe, other manual, semi-automatic, or automatic pump devices (e.g., a motorized pump) may also be utilized to carry out the sampling and purging operations disclosed herein, without departing from the broader aspects of the invention.

As discussed above, embodiments of the invention relate to pump devices and related methods to add or remove a known volume of fluid to or from a fluidic vessel in a functionally closed manner. The embodiments of the invention described herein are very simple and may be deployed as completely manual or as semi- or fully automated. In addition, the embodiments of the invention disclosed herein provide the ability to collect samples or make additions of almost any volume, in a repeatable and precise manner.

While the embodiments of the invention relate generally to pump devices and related methods to add or remove a known volume of fluid to or from a fluidic vessel in a functionally closed manner, the invention is not so limited in this regard, and it is contemplated that the inventive concepts disclosed herein may be applied to certain existing systems and devices to improve the functionality thereof. For example, the main chamber in the Sefia and Sepax devices from Cytiva is essentially a syringe barrel (that also capable of centrifugation), which can be utilized as the syringe in the embodiments of the invention disclosed herein. Moreover, for repeated sampling of a cell culture chamber, a variation of the inventive device (of suitable volume capacity) could be used either in true single-use fashion (a fresh device for every sample, for maximum reduction of contamination risk) or repeatedly for the duration of culture. The latter scenario would require the ability to flush the line of residue after each sampling event, such as by using the process disclosed above in connection withFIG.1.

In an embodiment, a sampling system is provided. The sampling system includes a graduated sampling chamber configured for fluid connection to a sample source, a pump device configured for fluid connection with the sampling chamber, and a sterile air filter intermediate the pump device and the sampling chamber, wherein the pump device is selectively actuatable to draw a volume of fluid from the sample source into the sampling chamber. In an embodiment, the sampling chamber includes a baffle separating an inlet, where the fluid enters the sampling chamber, from an outlet, where the pump draws air from the sampling chamber. In an embodiment, the system includes a first valve intermediate the sampling chamber and the sample source, the first valve permitting unidirectional flow of the fluid from the sample source to the sampling chamber. In an embodiment, the sampling chamber is configured for fluid connection to the sample source at a location adjacent to a bottom of the sampling chamber, the sampling chamber is configured for fluid connection to the pump device at a location adjacent to a top of the sampling chamber. In an embodiment, the system also includes a first valve intermediate the sampling chamber and the sample source, and a sample collection line fluidly connected to the sampling chamber via the first valve, wherein the first valve is actuatable to selectively place the sample source and/or the sample collection line in fluid communication with the sampling chamber. In an embodiment, the first value is movable to a first position where the sampling chamber is in fluid communication with the sample source, such that the pump device is operable draw the volume of fluid into the sampling chamber, and the first valve is movable to a second position where the sampling chamber is in fluid communication with the sample line so that the volume of fluid in the sampling chamber can flow from the sampling chamber through the sample collection line. In an embodiment, the pump device is a syringe. In an embodiment, the pump device is an automated pump. In an embodiment, the system includes a receptacle in fluid communication with the sample collection line. In an embodiment, the receptacle is a vacutainer. In an embodiment, the sample source is one of a cell culture vessel or a circulation loop. In an embodiment, the system further includes a second chamber configured for fluid connection to a media source via an inlet port and for fluid connection to the cell culture vessel or the circulation loop via an outlet port, a second pump device configured for fluid connection with the second chamber, a first valve intermediate the second chamber and the media source, the first valve permitting unidirectional flow from the media source to the second chamber, and a second valve intermediate the second chamber and the cell culture vessel or the circulation loop, the second valve permitting unidirectional flow from the second chamber to the cell culture vessel or the circulation loop. The second pump device is selectively actuatable to draw a volume of fluid from the media source into the second chamber, and to push the volume of fluid from the second chamber into the cell culture vessel or the circulation loop.

In another embodiment of the invention, a method for sampling is provided. The method includes the steps of connecting a sampling chamber to a sample source, and actuating a pump to draw a volume of fluid from the sample source through a valve, and into the sampling chamber, wherein the valve is configured to prevent backflow of fluid from the sampling chamber to the sample source. In an embodiment, the valve is one of a check valve or a stopcock. In an embodiment, the step of actuating the pump to draw the volume of fluid into the sampling chamber includes evacuating air from the sampling chamber through an outlet, wherein the outlet is configured with a sterile air filter. In an embodiment, the pump is a syringe, and the sampling chamber has graduated markings. In an embodiment, the method may also include the steps of opening a second valve to place a sampling line in fluid communication with the sampling chamber, and flowing the volume of fluid from the sampling chamber to the sampling line. In an embodiment, the step of flowing the volume of fluid from the sampling chamber to the sampling line includes pushing air into the sampling chamber through a sterile air filter to displace the volume of fluid from the sampling chamber.

In yet another embodiment, a bioprocessing system is provided. The bioprocessing system includes a cell culture vessel, and a first assembly for adding a first fluid to the cell culture vessel. The first assembly includes a first chamber configured for fluid connection to a source of the first fluid via an inlet port in the first chamber, and for fluid connection to the cell culture vessel via an outlet port in the first chamber, a first pump device configured for fluid connection with the first chamber, a first valve intermediate the first chamber and the source, the first valve permitting unidirectional flow from the source to the first chamber, and a second valve intermediate the first chamber and the cell culture vessel, the second valve permitting unidirectional flow from the first chamber to the cell culture vessel. The second pump device is selectively actuatable to draw a volume of the first fluid from the source into the first chamber, and to push the volume of fluid from the first chamber into the cell culture vessel. The bioprocessing system also includes second assembly for removing a second fluid from the cell culture vessel. The second assembly includes a second chamber configured for fluid connection to the cell culture vessel via an inlet port in the second chamber, and for fluid connection to a collection vessel via an outlet port in the second chamber, a second pump device configured for fluid connection with the second chamber, a third valve intermediate the cell culture vessel and the second chamber, the third valve permitting unidirectional flow from the cell culture vessel to the second chamber, and a fourth valve intermediate the second chamber and the collection vessel, the fourth valve permitting unidirectional flow from the second chamber to the collection vessel. The second pump device is selectively actuatable to draw a volume of the second fluid from the cell culture vessel into the second chamber, and to push the volume of fluid from the second chamber into the collection vessel. In an embodiment, the first pump and the second pump are syringes, and the first chamber and the second chamber have graduated markings.

As used herein, an element or step recited in the singular and proceeded with the word “a” or “an” should be understood as not excluding plural of said elements or steps, unless such exclusion is explicitly stated. Furthermore, references to “one embodiment” of the present invention are not intended to be interpreted as excluding the existence of additional embodiments that also incorporate the recited features. Moreover, unless explicitly stated to the contrary, embodiments “comprising,” “including,” or “having” an element or a plurality of elements having a particular property may include additional such elements not having that property.

This written description uses examples to disclose several embodiments of the invention, including the best mode, and also to enable one of ordinary skill in the art to practice the embodiments of invention, including making and using any devices or systems and performing any incorporated methods. The patentable scope of the invention is defined by the claims, and may include other examples that occur to one of ordinary skill in the art. Such other examples are intended to be within the scope of the claims if they have structural elements that do not differ from the literal language of the claims, or if they include equivalent structural elements with insubstantial differences from the literal languages of the claims.