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WO2025202193A1 - Système de tubulure pour transfert précis de liquides - Google Patents

Système de tubulure pour transfert précis de liquides

Info

Publication number
WO2025202193A1
WO2025202193A1 PCT/EP2025/058125 EP2025058125W WO2025202193A1 WO 2025202193 A1 WO2025202193 A1 WO 2025202193A1 EP 2025058125 W EP2025058125 W EP 2025058125W WO 2025202193 A1 WO2025202193 A1 WO 2025202193A1
Authority
WO
WIPO (PCT)
Prior art keywords
port
tubing
tubing system
liquid
channel
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
PCT/EP2025/058125
Other languages
English (en)
Inventor
Alexander UPSTONE
Daren Stephen Huttlestone
Adrian STACEY
Adrian Neil Bargh
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Automation Partnership Cambridge Ltd
Original Assignee
Automation Partnership Cambridge Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from EP24165955.6A external-priority patent/EP4624563A1/fr
Application filed by Automation Partnership Cambridge Ltd filed Critical Automation Partnership Cambridge Ltd
Publication of WO2025202193A1 publication Critical patent/WO2025202193A1/fr
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • 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
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/48—Automatic or computerized control
    • 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
    • C12M23/40—Manifolds; Distribution pieces
    • 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
    • C12M23/42—Integrated assemblies, e.g. cassettes or cartridges

Definitions

  • Bioprocesses of particular relevance for the present invention are in the area of cell and gene therapy, for example to manufacture autologous T cells that are modified to express a chimeric antigen receptor (CAR). These cells might be used for the treatment of various types of cancer, including different types of leukemia (blood cancer). Other cell therapies based on naive cells, in particular stem cells and their derivatives, are also of interest. Sterile and precise transfer of liquids is of utmost importance for the success of most bioprocesses, but often causes a number of technical problems. One issue arises when multiple pumps are involved, as each pump may have slight calibration differences, leading to flow variability and pressure differences at different points in the system, which ultimately leads to inaccurate volume transfers.
  • CAR chimeric antigen receptor
  • a significant challenge underlying the present invention is the presence of dead volume within the tubing and fittings.
  • Complex geometries, such as T-junctions or valve bodies, can trap liquid, leading to inaccurate dosing or loss of valuable product.
  • liquid may remain in sections of tubing that are not properly drained, while adsorption of proteins or other media components onto tubing surfaces can further reduce the effective volume transferred.
  • Maintaining sterility throughout the transfer process is also critical but difficult to ensure. Any break in sterility at connection points, such as sampling ports or sterile connectors, can introduce contamination risks. Small defects in tubing welds or connections may allow microbial ingress, while residual moisture within the system can promote biofilm formation. Furthermore, condensation inside the tubing may create an environment conducive to microbial growth if not properly managed.
  • the present invention addresses the issues outlined above by providing a novel tubing system designed for the controlled transfer of liquids in bioprocess applications.
  • This tubing system offers the advantage of enabling both bulk transfer and metered transfer between two ports, thereby enhancing operational flexibility.
  • the design of the tubing set significantly reduces the amount of dead volume, minimizing product loss and contamination risks.
  • tubing system Another key benefit of the tubing system is its ease of implementation.
  • the design allows for straightforward integration into existing bioprocess systems without the need for extensive modifications or specialized equipment. This simplicity not only reduces setup time and costs but also ensures reliable and consistent performance across various applications.
  • the tubing system according to the invention is particularly advantageous due to its one-size-fits-all design approach, allowing for a significant reduction in manufacturing costs compared to tubing systems tailored to a specific use case.
  • the invention provides a method for performing a bioprocess (also referred to as a bioprocessing method).
  • the method comprises: providing a bioprocessing station, providing a tubing system according to the first aspect, fluidically connecting the tubing system, which may be present in the form of a tubing cassette, to the bioprocessing station, preferably by welding, performing a method according to the second or third aspect, and disconnecting the tubing system from the bioprocessing station, preferably by sealing and cutting.
  • FIG. 3 a schematic representation of a third embodiment of a tubing system according to the first aspect of the invention
  • Fig. 8 a schematic illustration of a method for metered transfer of liquid
  • Fig. 9 a schematic illustration of a method for bulk transfer of liquid.
  • a tubing system comprising: a first port and a second port configured for connection to external elements; a main conduit fluidically connecting the first port and the second port; a valve associated with each port, each valve being operable independently to open or close the respective port, thereby regulating fluid flow along the main conduit; a metering vessel fluidically connected to the main conduit via a first channel, a valve being operable to open or close the metering vessel, thereby regulating fluid flow along the first channel; an air access point fluidically connected to the main conduit via a second channel, a valve being operable to open or close the air access point, thereby regulating fluid flow along the second channel; a bidirectional pump operatively connected to the main conduit; and pressure sensors located on opposite sides of the bidirectional pump.
  • the tubing system comprises at least two ports for connecting other elements of a bioprocess, such as tubing, reservoirs, bioreactors, media containers, waste vessels, additional tubing segments or the like.
  • the ports are implemented as standardized connectors that allow secure and sterile attachment of external elements.
  • These ports can take various forms depending on the specific requirements of the bioprocess, including sterility, ease of use, and compatibility with existing equipment.
  • Exemplary implementations of the ports are barbed connectors, Luer connectors, clamp fittings, flange connectors.
  • Preferred ports comprise a weldable first piece of sealed sterile tubing, such that another piece of weldable tubing can be welded to the first piece of tubing without breaking the sterile environment of the connector, particularly in cases where sterility must be maintained throughout a process.
  • ports may incorporate valved connectors, such as pinch valves, diaphragm seals, or quick-disconnect couplings with integrated shutoff features to prevent leaks when attaching or detaching components.
  • ports can also be designed for aseptic docking, where connections are made within sterile enclosures or using sterile welding techniques.
  • the tubing system is generally set up for sterile transfer of fluid between the ports or between pairs of ports.
  • the section of the tubing system between two ports is referred to as a “main conduit” in the present disclosure.
  • the transfer of fluid between the two ports generally ensues along the main conduit, although the fluid may be pumped at least intermittently along other parts of the tubing system, e.g. towards the metering vessel or away from it.
  • the tubing system comprises first and second channels, each sometimes referred to as a “side channel”, together “side channels”, throughout the present disclosure. These side channels fulfil important functions in the tubing system.
  • the first channel connecting the main conduit with the metering vessel enables use of the tubing system to perform precise metered transfer of liquids, as will be described elsewhere herein.
  • the second channel connecting the main conduit to the air access point serves as means for purging the tubing system of liquid.
  • Fluidical connections between any side channel(s) and/or main conduit(s) are referred to as “branching point” throughout the present disclosure.
  • Main conduit and side channels are together referred to as “channels” throughout the present disclosure.
  • the tubing system may comprise further ports for connecting additional external elements.
  • the maximum total number of ports a tubing system according to the present invention can accommodate is not limited, e.g. 3, 4, 5, 6, or even 10 or 15 ports or more are possible.
  • the tubing system further comprises at least one further port configured for connection to external elements, wherein each port is fluidically connected to every other port via a main conduit, wherein the main conduits can, independently from one another, be entirely separate or share a section of their length, and wherein at least one second channel opens into the main conduit at a location between each pair of ports, wherein preferably, a first channel leading to a metering vessel opens into the main conduit at a location between each pair of ports.
  • the tubing system is configured to facilitate more complex bioprocesses, such as, for example, a cell culture medium exchange procedure involving a cell culture medium vessel connected to a first port, a vessel for cultivating cells connected to a second port, and a waste vessel connected to a third port, wherein in a first step, spent cell culture medium may be moved from the second port to the third port, and in a subsequent second step, fresh cell culture medium may be moved from the first port to the second port.
  • complex bioprocesses such as, for example, a cell culture medium exchange procedure involving a cell culture medium vessel connected to a first port, a vessel for cultivating cells connected to a second port, and a waste vessel connected to a third port, wherein in a first step, spent cell culture medium may be moved from the second port to the third port, and in a subsequent second step, fresh cell culture medium may be moved from the first port to the second port.
  • the first port may be connected to the second port by a first conduit, wherein the liquid flow along the first conduit is regulated by a first set of valves, one being associated with the first port and one with the second port.
  • the first and second port may be connected by a second conduit, wherein the liquid flow along the second conduit is regulated by a second set of valves, one being associated with each of the ports.
  • the first conduit becomes the main conduit and liquid can flow though the first (main) conduit between the two ports.
  • the second conduit becomes the main conduit and liquid can flowthrough the second (main) conduit between the ports.
  • the tubing system achieves a liquid- tight connection between the external elements connected to the ports of the tubing system.
  • the term “fluidically connected” is intended to mean that one element of the tubing system is connected to another element of the tubing system so that fluid (i.e., liquid or gas) may flow between them without leaking; it is generally preferred that the only points of entry or discharge of fluids to/from the tubing system are the ports (for liquid and gas), and the air access point and optionally the metering vessel (each only for entry and discharge of gas). In this way, the tubing system ensures that a sterile environment is maintained within it.
  • the air access point (also referred to as “air inlet” throughout the present disclosure) may generally be designed as an open-ended piece of tubing, the end of which is covered by an air filter, such as a sterile filter having an average pore size of 220 nm or lower.
  • an air filter such as a sterile filter having an average pore size of 220 nm or lower.
  • At least one second channel may comprise a plurality of branch conduits adjacent to the main conduit, each branch conduit being in fluid communication with the main conduit at a distinct connection point, wherein each branch conduit comprises a valve being operable to open or close the respective branch conduit so as to allow fluid to flow independently through each branch conduit.
  • the connection points between the branch conduits and the main conduit may be placed on both sides of a pump located in the main conduit or in a first channel connecting to the main conduit between the connection points of the branch conduits. In this arrangement, the entire tubing system can be effectively purged of liquid without leaving any dead volume in the system, which is advantageous when pumping high- value liquid or, e.g., when aiming for precise metered transfer of liquid.
  • the tubing may be produced from any material known to be compatible with bioprocessing, including but not limited to silicone, thermoplastic elastomers (TPEs), polyvinyl chloride (PVC), fluoropolymers such as PTFE, polyethylene (PE), polypropylene (PP) or ethylene vinyl acetate (EVA).
  • TPEs thermoplastic elastomers
  • PVC polyvinyl chloride
  • fluoropolymers such as PTFE
  • PE polyethylene
  • PP polypropylene
  • EVA ethylene vinyl acetate
  • the material choice may depend on factors like fluid type, sterility requirements, biocompatibility, and resistance to pressure, temperature, and gas exchange. The skilled person is capable of choosing the material to achieve a balance of chemical compatibility, flexibility, durability, and sterility.
  • the tubing system comprises a metering vessel.
  • the metering vessel is configured to define a controllable volume within the tubing system, allowing for precise measurement and transfer of liquid. Its structure is tailored to ensure compatibility with the overall fluid pathway while maintaining sterility and minimizing dead volume.
  • the metering vessel is integrated into the tubing system in such a way that it can be filled and emptied in a controlled manner, e.g. by the action of a pump.
  • the metering vessel may incorporate flexible or rigid walls, depending on the intended mode of operation.
  • a flexible-walled vessel for instance, can be compressed to expel liquid or expanded to draw liquid in, functioning similarly to a diaphragm-based dosing chamber.
  • a rigid-walled metering vessel may rely on precisely controlled inflow and outflow through associated valve mechanisms.
  • the vessel may also feature volume markings or be paired with external sensors to facilitate monitoring of the liquid level.
  • the metering vessel comprises an air inlet connected to it to allow for inflow and outflow of sterile air, thus avoiding overpressurisation or creation of a vacuum.
  • the air inlet may be in the form of an open-ended piece of tubing having a sterile air filter attached at its end.
  • the air inlet may be arranged in a top part of the metering vessel, such that it is not contacted by liquid flowing into the metering vessel.
  • the metering vessel is generally arranged inline within a first channel of the tubing system, which connects the metering vessel to the main conduit.
  • the metering vessel is accessed by opening/closing a valve, which is preferably one that has no dead volume.
  • the metering vessel is positioned at the end of the first channel within the tubing system, such that it has a single connection point through which liquid is both received and dispensed. Unlike other components of the tubing system, which may feature multiple connection points to accommodate different flow paths, the metering vessel interfaces with the system through a single inlet, ensuring controlled fluid transfer. In systems requiring highly accurate dosing, the vessel may be designed to accommodate a defined liquid volume with minimal expansion or contraction, ensuring repeatability in metered transfers. Regardless of its specific geometry, the metering vessel serves as a critical element in achieving controlled liquid handling while maintaining sterility and compatibility with the single-use nature of the tubing system.
  • the metering vessel is configured for measurement of the liquid volume present therein. This is preferably achieved by the metering vessel being connected to a weighing system. Therein, the measurement is preferred to be a continuous and/or real-time measurement.
  • the metering vessel (MV) is configured for being reversibly sealed and unsealed, wherein preferably, reversible sealing is achieved through welding.
  • the ability to weld and seal/cutthe metering vessel for reversibly connecting to the tubing system enables secure and sterile integration of the metering vessel into the system while allowing for its removal without compromising system integrity. Welding ensures a seamless and leak-free connection, reducing the risk of contamination and maintaining sterility throughout operation.
  • the option to seal and cut the vessel further allows for controlled disconnection, enabling the option to replace the metering vessel as a consumable component without exposing the tubing system to environmental contaminants and without having to replace the entire tubing system when performing multiple consecutive metered transfers.
  • the metering vessel may be used to acquire an aliquot of the pumped liquid without compromising the sterile environment within the tubing system by pumping a metered amount of liquid into the metering vessel, performing sealing and cutting, and removing the metering vessel from the tubing system, e.g. for further analysis or storage of the aliquot.
  • the tubing system comprises a bidirectional pump for pumping fluid through the channels and conduits of the tubing system.
  • the term “bidirectional” is intended to infer the usual meaning of the term, which is that the pump can be controlled/actuated to pump fluid into a first direction, and the same pump can alternatively be controlled/actuated to pump the fluid into a second direction opposite the first direction.
  • the tubing system is generally compatible with encompassing more than one pump, arrangements including multiple pumps have certain disadvantages as alluded to above. It is therefore generally preferred that the tubing system comprises exactly one pump.
  • the bidirectional pump is a peristaltic pump.
  • peristaltic pumps consist of a pump head comprising a central rotating mechanism with multiple rollers or shoes for sequentially compressing the adjacent tubing.
  • peristaltic pumps further comprise an occlusion track, which is typically a curved surface against which the tubing is compressed by the rollers.
  • the rotating rollers squeeze the tubing against the occlusion track, pinching it completely closed at the point of contact. This prevents backflow and isolates discrete segments of fluid.
  • each roller moves along the tubing, pushing the fluid trapped between successive points of occlusion forward.
  • peristaltic pumps have the advantage that the pump head can be provided as a reusable pre-installed component of, for example, a bioprocessing station, and can be used in combination with a consumable tube set which is fitted to the pump head when arranged on the bioprocessing station, e.g. in the form of a millifluidic module.
  • the pump is operatively connected to the main conduit.
  • the pump is functionally integrated with the conduit in such a way that it can actively influence fluid flow within the main conduit.
  • This does not necessarily mean a direct physical attachment to the main conduit - rather, it signifies that the pump interacts with the tubing system to move, regulate, or control the fluid inside the main conduit.
  • the location of the pump within the tubing system is important for the pumping functionality of the tubing system. For example, if bulk liquid transfer is intended, it is necessary to arrange the pump within the main conduit. In this configuration, the pump can directly act onto the main conduit to pump fluid from one port through the main conduit directly to another port, i.e. without pumping into the metering vessel. Alternatively, for example, if metered transfer is intended, the pump may be arranged along a side channel, e.g. along the first channel, i.e. between the branching point and the metering vessel, or between the main conduit and the metering vessel.
  • a bidirectional pump is located between two branch conduits of a second channel leading to an air access point.
  • This configuration allows air to be introduced from both sides of the pump, effectively displacing any remaining liquid and eliminating dead volume within the system.
  • this design optimizes liquid recovery, which is particularly advantageous when handling high-value liquids or when precise metered transfer is required.
  • the ability to fully purge the system further enhances accuracy and reproducibility, while also minimizing product loss and preventing crosscontamination between process steps.
  • the first channel and the second channel are configured to connect to the main conduit via a unified connection by sharing at least a section of their length, preferably adjacent to the main conduit.
  • the bidirectional pump is located in the shared section of the first channel and the second channel. This arrangement simplifies the tubing system by reducing the number of pumps required, minimizing mechanical complexity and potential failure points. With both liquid and air traveling through the same pump, transitions between liquid transfer and system purging can be managed efficiently without the need for additional valving. This design is particularly advantageous in compact setups where space constraints or system simplification are priorities, while still ensuring effective purging and controlled metered transfer.
  • a first channel opens into the main conduit at a location between each pair of ports. This configuration is particularly advantageous in tubing systems comprising more than two ports, as it provides for metered transfer of liquids between each pair of ports.
  • the tubing system incorporates two pressure sensors which are positioned on either side of the bidirectional pump, enabling differential pressure monitoring across the pumping mechanism.
  • This arrangement allows for a more precise assessment of pump performance by detecting pressure drops, ensuring consistent flow rates, and identifying variations that may result from system irregularities such as filter clogging, tubing deformation, or air bubble formation.
  • the system can dynamically adjust operating parameters to maintain optimal liquid transfer conditions.
  • the use of two pressure sensors further enhances process control by enabling closed-loop feedback regulation, where real-time pressure data can be used to adjust pump speed or valve operation in response to changing conditions.
  • the tubing system comprises at least one pressure sensor associated with each further pump, ideally two pressure sensors located on either side of each further pump.
  • the main conduits between all ports of the tubing system share a section of their length. This section of tubing is also referred to herein as the “shared main conduit”.
  • the bidirectional pump is located in the shared main conduit, thereby enabling the system to perform bulk transfer between each pair of ports.
  • the first channel leading to the metering vessel may open into the shared main conduit, enabling the possibility of metered transfer between each pair of ports.
  • at least one second channel, preferably all second channels, most preferably all first and second channels open(s) into the shared main conduit.
  • the shared main conduit may comprise the following branching points in succession: a second channel leading to an air access point, the bidirectional pump, a first channel leading to the metering vessel, and another second channel leading to the same or a different air access point.
  • first port and the second port shall not be limited to this specific combination of ports, but shall encompass transfer between any combination of two ports of the tubing system (e.g., from the second port to the first port, or from the first or second port to a further port).
  • the present disclosure only refers to first and second port in order to remain easy to understand.
  • a defined amount (first amount) of a liquid is transferred from a first port into a metering vessel.
  • the liquid passes through the following consecutive elements of the tubing system: a valve associated with the first port, optionally a section of the main conduit connecting the first port with another port, at least a section of the first channel, and the valve associated with the metering vessel.
  • the liquid passes through a pump, preferably through the bidirectional pump, which may be located in the main conduit (e.g., in the shared main conduit) or in the first channel.
  • the bidirectional pump is controlled to pump fluid towards the metering vessel until it is determined that the first amount of liquid has been transferred. This may be achieved by any means known in the art, for example by determining the weight of the metering vessel and causing stopping of the pump when a predetermined weight difference of the metering vessel is observed.
  • the tubing section between the first port and the metering vessel is cleared from the liquid.
  • the liquid is cleared by transferring the remaining liquid to the first port, whereby unused liquid is preserved and does not need to be discarded or otherwise removed from the system.
  • the clearance is generally achieved using the air inlet, e.g. by opening the valves associated with the air access point and the first port and pumping towards the first port, thereby replacing the liquid with air from the air inlet, preferably until the entire distance between air access port and first port contains only air.
  • the pumping should continue until the remaining liquid has at least passed through the valve associated with the first port.
  • the section cleared may be the entire tubing between the first port and the metering vessel, e.g. in case that the metering vessel is directly connected to a main conduit or second channel (i.e. if the first channel has no volume).
  • the section cleared can also be a defined section between first port and metering vessel, e.g. the section between first port and branching point of the first channel.
  • the metered volume includes the amount of liquid present in the uncleared section of tubing, which can e.g. correspond to the volume of the first channel.
  • a metered amount of the liquid is pumped from the metering vessel to a second port.
  • This is preferably achieved by pumping through the bidirectional pump, e.g. in a tubing system where the bidirectional pump acts on the side channel or main conduit between the metering vessel and the second port.
  • the pumping is generally performed until the entire metered amount has passed a fixed point of reference, which may be close to the metering vessel.
  • the passing of the metered amount can be detected, for example, by continuously determining the weight of the metering vessel and comparing the determined amount with a target weight of the metering vessel which is representative of the metered amount having passed the fixed point of reference.
  • the fixed point of reference may be a branching point of the first channel leading to the metering vessel with a second channel or a main conduit.
  • the metered amount can have the same volume as the first amount pumped into the metering vessel (which is possible but less preferred), or can have a lower volume, e.g. the first amount minus a predetermined dead volume which remains as a residue inside the metering vessel or first channel (which is preferred).
  • a lower volume e.g. the first amount minus a predetermined dead volume which remains as a residue inside the metering vessel or first channel (which is preferred).
  • the latter approach enables rapid filling of the metering vessel, followed by controlled dispensing at a reduced flow rate to achieve precise volumetric delivery.
  • the metering vessel may have a variable volume (e.g. by being implemented as a flexible container) and/or may preferably have a separate air inlet connected to it, thus also avoiding high vacuum pressure.
  • the section cleared may be the entire tubing between the metering vessel and the second port, e.g. in case that the metering vessel is directly connected to a main conduit or second channel (i.e. if the first channel has no volume).
  • the section cleared can also be a defined section between metering vessel and second port, e.g. the section between the branching point of the first channel and the second port.
  • the metered volume does not include the amount of liquid present in the uncleared section of tubing, which can e.g. correspond to the volume of the first channel.
  • the method according to the second aspect may involve clearing the metering vessel and the first channel from the remaining liquid.
  • This can be implemented by transferring the remaining liquid to the first port in order to recover non-transferred volume retained there.
  • all liquid is removed from the tubing system, and any liquid not transferred is preserved for later use.
  • the remaining liquid can be transferred to a waste outlet, e.g. to another port connected to a waste container, thereby directly discarding any remaining liquid not intended for recovery, such as a transfection mixture.
  • This step can be achieved, for example, by pumping from the metering vessel to the first port or waste outlet, thereby replacing the liquid with air drawn in through an air inlet of the metering vessel.
  • the method according to the second aspect of the present invention is particularly advantageous because it allows for high precision of the transferred volume. This is ensured by having the pump cleared from metered liquid from both sides: once from the side opposite the first port after pumping the liquid to the metering vessel, and once from the side opposite the second port after pumping the liquid towards the second port.
  • the method according to the third aspect is intended to transfer a large amount of liquid directly between two external elements connected to two ports of the tubing system according to the first aspect. It is not necessary to precisely determine or measure the amount transferred; it is, however, possible to do so, e.g. by weighing one or both of the external elements and causing stopping of the transfer once a predetermined weight difference is measured. Another exemplary possibility is measuring the pressure inside the tubing system as described elsewhere herein and causing stopping of the transfer once the pressure exceeds or falls below a limit value.
  • the method comprises, in a first step, transferring an intended bulk amount of the liquid from a first port into a second port through a bidirectional pump.
  • the bulk amount of liquid passes through at least the following consecutive components of the tubing system according to the first aspect: the first port, the valve associated with the first port, a main conduit connecting the first port with the second port, the bidirectional pump, the valve associated with the second port, and the second port.
  • This step ensues until the transfer is caused to stop, e.g. by actuating the bidirectional pump to stop pumping towards the second port.
  • the method according to the third aspect comprises clearing the tubing section between the bidirectional pump and the second port by transferring the remaining liquid to the second port, and/or clearing the tubing section between the first port and the bidirectional pump by transferring the remaining liquid to the first port, e.g. using air drawn through the air inlet or through the metering vessel.
  • the term “remaining liquid” refers to the part of the bulk liquid which is present in the referred-to part of the tubing system, i.e. the liquid which has exited the first port and travelled through the main conduit to the bidirectional pump.
  • this can be achieved by having an air inlet arranged on the side of the bidirectional pump opposite the second port (along the conduits and channels), opening the valves associated with the air inlet and the second port, closing all other valves, and pumping towards the second port.
  • This step ensures that all liquid exits the tubing system, thereby reducing the amount of waste produced by the system.
  • this can be achieved by having an air inlet arranged on the side of the bidirectional pump opposite the first port (along the conduits and channels), opening the valves associated with the air inlet and the first port, closing all other valves, and pumping towards the second port.
  • the method according to the invention allows for complete clearance of the tubing system from liquid. This effect is of particularly advantageous when transferring high-value liquids, as it ensures that no liquid is wasted in the course of transferring through the tubing system according to the invention.
  • the method according to this exemplary embodiment comprises the following steps: transferring a priming amount of the liquid from a first port to a metering vessel transferring an intended bulk amount of the liquid from the first port into a second port through a bidirectional pump, clearing the tubing section between the bidirectional pump and the second port by transferring the remaining liquid to the second port, and clearing the tubing section between the first port and the metering vessel by transferring the remaining liquid to the first port using the metering vessel.
  • the priming amount of liquid has at least the same volume as the tubing between the first port and the metering vessel.
  • the steps of clearing the tubing sections between the pump and the respective ports can be performed in reverse order without having to further adapt the method in any way, i.e. the method according to the third aspect encompasses an embodiment wherein the tubing section between the bidirectional pump and the first port is cleared first, and afterwards the tubing section between the bidirectional pump and the second port is cleared.
  • a tubing cassette comprising a tubing system according to the first aspect of the present invention.
  • the tubing cassette is generally configured that the tubing system contained therein can be fluidically connected to an external element in a reversible fashion.
  • cassette is intended to refer to a structured interface for integrating the tubing system into a broader process environment while providing a controlled and organized layout for fluid transfer.
  • the cassette typically defines a firm surface that defines connection points for external elements and other components such as sensors or pumps, enabling secure integration and controlled fluid handling.
  • the cassette enhances ease of use, facilitates reliable and sterile connections, and contributes to overall process efficiency by reducing setup complexity and ensuring consistent performance.
  • the membrane serves as an integral part of the valve mechanism, as it can be selectively deformed into the recessed areas to block fluid flow through the corresponding channels, thereby functioning as a valve.
  • a second molded body may be positioned on the opposite side of the membrane, providing structural support while incorporating through-holes aligned with the valve regions. These through-holes allow actuation elements, such as pistons, to extend through the second molded body and press against the membrane at designated points, selectively controlling fluid passage within the channels. This arrangement enables precise and reliable valve operation while maintaining a compact and enclosed design that facilitates integration into a broader fluid handling system.
  • a bioprocessing method comprising: providing a bioprocessing station, providing a tubing system according to the first aspect of the present invention, fluidically connecting the tubing cassette to the bioprocessing station, preferably by welding, performing a method according to the second or third aspect of the present invention, and disconnecting the tubing system from the bioprocessing station, preferably by sealing and cutting.
  • the bioprocessing method comprises the use of the tubing system according to the first aspect of the invention and a bioprocessing station configured to interact with the tubing system during operation.
  • the method includes providing the tubing system in a state suitable for fluid handling, wherein the tubing system comprises a structured arrangement of channels and connection points for interfacing with external elements and components of the bioprocessing station such as valve actuators.
  • the bioprocessing module is arranged to facilitate controlled fluid transfer through the tubing system, enabling the execution of process steps necessary for the intended bioprocessing application.
  • the bidirectional pump of the tubing system may be implemented in that the bioprocessing station comprises a peristaltic pump head and the tubing system comprises a protruding tubing section aligning and mating with the peristaltic pump head.
  • the consumable part of the tubing system comprises no electronic components and is therefore easier to mass-produce and discard.
  • the bioprocessing method further includes performing a bioprocess, wherein fluid is directed through the tubing system in accordance with a defined sequence of operations as laid out above in the context of the second and third aspects.
  • the interaction between the tubing system and the bioprocessing station ensures that process conditions are maintained within predefined parameters, supporting the controlled progression of the bioprocess.
  • the tubing system is disconnected from the bioprocessing station. Disconnection may involve sealing and detaching fluid connections, removing physical interfaces, and/or deactivating integrated control mechanisms.
  • the disconnection process is preferably performed in a manner that maintains the integrity of both the tubing system and the bioprocessing module, ensuring that any remaining fluid is managed appropriately and that system components remain in a state suitable for subsequent use or disposal.
  • bioprocesses according to the present invention include, but are not limited to, the following:
  • the tubing system is present in the form of a cassette according to the fourth aspect of the invention.
  • the method enables reliable, repeatable, and scalable bioprocessing while allowing for the efficient exchange of system components.
  • the ability to disconnect the cassette upon process completion facilitates ease of operation, supports sterility management, and enhances process flexibility by allowing for rapid system reconfiguration or replacement.
  • Embodiments referencing others also refer to previous embodiments numbers containing a letter. For example, “Tubing system according to embodiment 1 to 5” also reverts back to embodiment 4a.
  • Tubing system comprising: a first port (T2) and a second port (T4) configured for connection to external elements; a main conduit fluidically connecting the first port (T2) and the second port (T4); a valve (V10, V8) associated with each port (T2, T4), each valve (V10, V8) being operable independently to open or close the respective port (T2, T4), thereby regulating fluid flow along the main conduit; a metering vessel (MV) fluidically connected to the main conduit via a first channel, a valve (V6) being operable to open or close the metering vessel (MV), thereby regulating fluid flow along the first channel; an air access point (F) fluidically connected to the main conduit via a second channel, a valve (V7) being operable to open or close the air access point (F), thereby regulating fluid flow along the second channel; a bidirectional pump operatively connected to the main conduit; and pressure sensors located on opposite sides of the bidirectional pump.
  • Tubing system according to embodiment 1 wherein the metering vessel (MV) is configured for measurement of the fluid volume present therein. .
  • Tubing system according to embodiment 2 wherein the metering vessel (MV) is connected to a weighing system.
  • Tubing system according to embodiment 2 or 3 wherein the measurement is a continuous and/or real-time measurement.
  • a. Tubing system according to any one of the preceding embodiments, wherein the metering vessel comprises an air inlet connected to it.
  • the air inlet is in the form of an open-ended piece of tubing having a sterile air filter attached at its end.
  • each of the first and second channels comprises a separate bidirectional pump.
  • each port (Tl, T2, T3, T4] is fluidically connected to every other port via a main conduit.
  • Tubing system according to any one of embodiments 24 to 26, wherein in addition to two second channels, a first channel opens into the main conduit at a location between each pair of ports.
  • Method for transferring a metered amount of a liquid using a tubing system comprising: transferring a first amount of the liquid from a first port into a metering vessel, preferably through a bidirectional pump, clearing the tubing section between the first port and the metering vessel from the liquid by transferring the remaining liquid to the first port using the air access point, pumping a metered amount of the liquid from the metering vessel to a second port, preferably through the bidirectional pump, and clearing the tubing section between the metering vessel and the second port from the liquid by transferring the remaining liquid to the second port using the air access point.
  • Method according to embodiment 28, further comprising: clearing the metering vessel and the first channel from remaining liquid.
  • Method according to embodiment 28a wherein the remaining liquid is transferred to the first port.
  • Method according to embodiment 28a. wherein the remaining liquid is transferred to a waste outlet.
  • Method for transferring a bulk amount of a liquid using a tubing system according to any one of embodiments 17 to 21 or 24 to 27, the method comprising: transferring an intended bulk amount of the liquid from a first port into a second port through a bidirectional pump, clearing the tubing section between the bidirectional pump and the second port by transferring the remaining liquid to the second port, and/or clearing the tubing section between the first port and the bidirectional pump by transferring the remaining liquid to the first port, e.g.
  • Method according to embodiment 29, comprising: transferring a priming amount of the liquid from a first port to a metering vessel transferring an intended bulk amount of the liquid from the first port into a second port through a bidirectional pump, clearing the tubing section between the bidirectional pump and the second port by transferring the remaining liquid to the second port, and clearing the tubing section between the first port and the metering vessel by transferring the remaining liquid to the first port using the metering vessel.
  • Tubing cassette comprising a tubing system according to any one of embodiments 1 to 27, wherein the tubing cassette is configured that the tubing system contained therein can be fluidically connected to an external element in a reversible fashion.
  • Tubing cassette according to any one of embodiments 35 to 37, wherein the second molded body comprises sensor openings positioned to correspond with pressure sensor regions of the flexible membrane.
  • Tubing cassette according to embodiment 30, comprising a rigid housing enclosing a flexible tubing set.
  • Bioprocessing method comprising: providing a bioprocessing station, providing a tubing system according to any one of embodiments 1 to 27, fluidically connecting the tubing system to the bioprocessing station, preferably by welding, performing a method according to embodiment 28 or 29, and disconnecting the tubing system from the bioprocessing station, preferably by sealing and cutting.
  • Fig- 1 shows a schematic illustration of a minimal embodiment of a tubing system according to the present invention.
  • the tubing system 1 comprises a first port T2 and a second port T4.
  • the ports T2, T4 are configured to connect to external elements.
  • the ports function as entry and exit points of fluid to be transferred through the tubing system 1.
  • Associated with the first and second port T2, T4 are valves V2 and V4, respectively.
  • the valves V2, V4 can be opened and closed independently of one another, e.g. by being independently actuatable to open or close.
  • the tubing between the ports T2, T4 is the main conduit 2.
  • a side channel 4 opens into the main conduit 2.
  • the side channel 4 is a common section of the first channel 5 leading to the metering vessel MV and the second channel 6 leading to the air access point F.
  • the metering vessel MV and air access point F are operable to open or close by their associated valves V6 and V7, respectively. These valves regulate the flow of fluid along their respective channel.
  • the bidirectional pump P is located in the common section of the side channel 4, thereby operatively connected to the main conduit 2.
  • the pump P is flanked by two pressure sensors PSI, PS2, which connect to the side channel 4 at branching points on either side of the pump P.
  • PSI pressure sensors
  • the pump P is implemented as a section of tubing extending between the connection points Pla and Plb which can interface with a peristaltic pump head able to facilitate bidirectional pumping.
  • valve V6 is closed and valve V7 is opened, and the pump P is controlled to pump fluid towards the first port T2 through the valve V2 which remained open.
  • This pumping is continued until the transferred liquid has been replaced by air in the entire section between the metering vessel MV and the first port T2, which can be confirmed visually, or by measuring the pressure e.g. with one or both of the pressure sensors PSI, PS2, or e.g. by reading out signals from a liquid presence sensor.
  • valves V2 and V7 are closed, valves V4 and V6 are opened, and the pump P is controlled to pump towards the second port T4. This pumping is continued until the metered amount of liquid has exited the metering vessel MV, which can be confirmed, e.g., by determining the weight of the metering vessel MV.
  • the part of the metered amount remaining in the tubing system 1, i.e. the part that has not yet entered the second port T4, must be cleared from the tubing system 1. This is achieved by closing valve V6 and opening valve V7 and continuing to pump fluid towards the second port T4. This pumping may continue, for example, until the transferred liquid has been replaced by air in the entire section between the metering vessel MV and the second port T4, which can be confirmed visually, or by measuring the pressure e.g. with one or both of the pressure sensors PSI, PS2, or e.g. by reading out signals from a liquid presence sensor. In an optional final step, the liquid remaining in the metering vessel MV can be recovered from the tubing system.
  • valves V7 and V4 are closed and valves V2 and V6 are opened.
  • the bidirectional pump P is controlled to pump towards the first port T2, i.e. towards valve V2, thereby replacing the liquid remaining in the metering vessel MV and in the first channel 5 with air entering the metering vessel MV through an air inlet.
  • This pumping may continue, for example, until the transferred liquid has been replaced by air in the entire section between the metering vessel MV and the first port T2, which can be confirmed visually, or by measuring the pressure e.g. with one or both of the pressure sensors PSI, PS2, or e.g. by reading out signals from a liquid presence sensor.
  • Fig- 2 shows an illustration of another embodiment of a tubing system 1 according to the present invention.
  • This embodiment differs from the one shown in Fig. 1 by the added valves V10 and V8 which are associated with the first and second ports, respectively.
  • the tubing system 1 allows for metered transfer according to the second aspect of the present invention as well as for bulk transfer according to the third aspect of the present invention.
  • Metered transfer is accomplished as described in relation to Figure 1.
  • a source element e.g., a container filled with a liquid
  • a destination element e.g., an empty container
  • valves V2 and V6 are opened, and the pump P is controlled to pump towards the metering vessel MV, i.e. towards valve V6. This pumping is continued until a priming amount of liquid has been transferred towards the metering vessel. Including this step has the effect that dead volume effects within the tubing system 1 are reduced and the liquid can be transferred with improved accuracy.
  • valve V6 is closed and valve V8 is opened, with the other valves being closed, and the pump P is controlled to pump towards the second port (i.e., towards valve V8) along the main conduit 2.
  • This pumping is continued until the bulk amount of liquid has been transferred to the second port T4.
  • valve V8 is closed and valve V7 associated with the air access point is opened.
  • the pump P is controlled to pump towards the first port (i.e., towards valve V2, which remains open). This pumping is continued until no liquid remains between the bidirectional pump and the first port.
  • valves V10 and V4 are opened instead of valves V2 and V8 before initiating the bulk transfer, and the pump P is controlled to pump in the opposite direction (i.e., towards valve V4).
  • valve V10 is closed and valve V7 is opened, and the pump P is controlled to pump towards the second port (i.e., towards valve V4) until the section between the bidirectional pump and the second port is cleared of liquid.
  • valves V2 and V8 are opened, with the other valves being closed, and the pump P is controlled to pump towards the second port (i.e., towards valve V8) along the main conduit 2. This pumping is continued until the bulk amount of liquid has been transferred to the second port T4.
  • valve V8 is closed and valve V7 associated with the air access point is opened.
  • the pump P is controlled to pump towards the first port (i.e., towards valve V2, which remains open). This pumping is continued until no liquid remains between the bidirectional pump and the first port.
  • Fig. 4 shows another embodiment of the tubing system according to the first aspect of the invention. This embodiment corresponds to the one shown in Fig. 3, except that it contains two additional ports, resulting in a total of four ports (Tl, T2, T3, T4).
  • Each port is associated with at least one valve regulating fluid flow along a respective channel: port Tl is associated with valve VI, port T2 is associated with valves V2 and V10, port T3 is associated with valves V3 and V9, and port T4 is associated with valves V4 and V8.

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Abstract

La présente invention concerne un système de tubulure destiné au transfert contrôlé de liquides dans un bioprocédé. L'invention concerne en outre, à l'aide du système de tubulure, un procédé de transfert d'une quantité mesurée d'un liquide et un procédé de transfert d'une quantité en vrac d'un liquide. L'invention concerne en outre une cassette de tubulure contenant le système de tubulure. L'invention concerne en outre un procédé de biotraitement impliquant le système de tubulure.
PCT/EP2025/058125 2024-03-25 2025-03-25 Système de tubulure pour transfert précis de liquides Pending WO2025202193A1 (fr)

Applications Claiming Priority (14)

Application Number Priority Date Filing Date Title
EP24165955.6A EP4624563A1 (fr) 2024-03-25 2024-03-25 Procédé pour effectuer un bioprocédé sur des cultures cellulaires immunitaires ou naïves
EP24165955.6 2024-03-25
EP24214283.4 2024-11-20
EP24214283 2024-11-20
EP24214275 2024-11-20
EP24214265 2024-11-20
EP24214265.1 2024-11-20
EP24214275.0 2024-11-20
EP24222584 2024-12-20
EP24222584.5 2024-12-20
EP24222573.8 2024-12-20
EP24222573 2024-12-20
EP25152692 2025-01-17
EP25152692.7 2025-01-17

Publications (1)

Publication Number Publication Date
WO2025202193A1 true WO2025202193A1 (fr) 2025-10-02

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Application Number Title Priority Date Filing Date
PCT/EP2025/058146 Pending WO2025202206A1 (fr) 2024-03-25 2025-03-25 Procédé de réalisation d'un bioprocédé sur des cultures de cellules immunitaires ou naïves
PCT/EP2025/058125 Pending WO2025202193A1 (fr) 2024-03-25 2025-03-25 Système de tubulure pour transfert précis de liquides
PCT/EP2025/058073 Pending WO2025202170A1 (fr) 2024-03-25 2025-03-25 Module de biotraitement mobile, unité de biotraitement et système de biotraitement pour la mise en œuvre d'un bioprocédé
PCT/EP2025/058145 Pending WO2025202205A1 (fr) 2024-03-25 2025-03-25 Base de biotraitement, station de biotraitement et système de biotraitement
PCT/EP2025/058149 Pending WO2025202209A1 (fr) 2024-03-25 2025-03-25 Procédé de mise en œuvre d'un bioprocédé sur une ou plusieurs cultures de cellules immunitaires ou naïves
PCT/EP2025/058162 Pending WO2025202218A1 (fr) 2024-03-25 2025-03-25 Récipient pour la culture de cellules dans un milieu liquide dans une procédure de culture
PCT/EP2025/058159 Pending WO2025202216A1 (fr) 2024-03-25 2025-03-25 Procédé de réalisation d'un bioprocédé sur des cellules immunitaires ou naïves dans une culture cellulaire

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Application Number Title Priority Date Filing Date
PCT/EP2025/058073 Pending WO2025202170A1 (fr) 2024-03-25 2025-03-25 Module de biotraitement mobile, unité de biotraitement et système de biotraitement pour la mise en œuvre d'un bioprocédé
PCT/EP2025/058145 Pending WO2025202205A1 (fr) 2024-03-25 2025-03-25 Base de biotraitement, station de biotraitement et système de biotraitement
PCT/EP2025/058149 Pending WO2025202209A1 (fr) 2024-03-25 2025-03-25 Procédé de mise en œuvre d'un bioprocédé sur une ou plusieurs cultures de cellules immunitaires ou naïves
PCT/EP2025/058162 Pending WO2025202218A1 (fr) 2024-03-25 2025-03-25 Récipient pour la culture de cellules dans un milieu liquide dans une procédure de culture
PCT/EP2025/058159 Pending WO2025202216A1 (fr) 2024-03-25 2025-03-25 Procédé de réalisation d'un bioprocédé sur des cellules immunitaires ou naïves dans une culture cellulaire

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WO2025202206A1 (fr) 2025-10-02
WO2025202170A1 (fr) 2025-10-02
WO2025202218A1 (fr) 2025-10-02
WO2025202216A1 (fr) 2025-10-02
WO2025202205A1 (fr) 2025-10-02

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