Liquid sampling device

Abstract

A device for sterile, small volume sampling of liquids includes a flow control system and a piping manifold. The conduit manifold includes an air inlet, a sample inlet, an air outlet, and a sample outlet, each of the air inlet, the sample inlet, the air outlet, and the sample outlet being fluidly coupled to the conduit manifold and operably coupled to the flow control system. A flow control system (FCS) is operable in a downstream configuration to pump a predetermined volume of fluid from a sample inlet to a sample outlet. After pumping the predetermined volume of fluid, the flow control system is operable in a countercurrent configuration to return fluid remaining in the conduit manifold to the sample inlet. The down-flow configuration and the counter-flow configuration substantially prevent the formation of bubbles in the fluid exiting the sample outlet or the sample inlet.

Description

Technical Field

[0001] Generally speaking, the present invention relates to a device for aseptic small volume sampling of liquids. In particular, the present invention relates to, but is not limited to, aseptic small volume sampling, wherein the main fluid can be returned to the main fluid reservoir while maintaining the sterility of the main fluid. Background Art

[0002] Current cell sampling methods require manual and open manipulation of cell cultures. This can introduce contaminants. However, sampling is a critical step often performed in bioprocessing and cell manufacturing because cells are monitored via off-line assays. Non-invasive on-line monitoring is not a standard practice for these industries.

[0003] Traditional solutions for cell sampling are to use aseptic techniques performed in a Biosafety Cabinet (BSC). Using aseptic techniques, the cell culture container is opened in the BSC so that a sample can be removed using a micropipette or a serum pipette. However, open manipulation presents an opportunity for contamination and relies on the aseptic technique of a trained operator; manual handling requires the full attention of a trained operator; using a BSC incurs indirect costs and time; access to the cell culture is required and this is not compatible with a closed automated system; and moving the culture between the incubator and the BSC disrupts the optimal conditions for the culture.

[0004] Another method involves sampling via needleless swabbable valves. The needleless connector is wiped with an alcohol wet wipe and connected to create a fluid path to extract a sample volume. However, the tubing preserves a dead volume of the sample and this volume must be flushed out before obtaining the next sample. This wastes cell samples because the cell culture left in the tubing cannot be returned to the cell culture aseptically; the sample previously left in the tube may cause inaccurate future cell sampling / monitoring due to sample damage and decay; and there are concerns about contamination via the swabbable valve.

[0005] There are also many other methods, each having its own drawbacks in terms of the opportunity for contamination and waste of cell cultures. Therefore, it is desirable to develop a device or system that avoids or ameliorates more than one of the above drawbacks of the existing methods, or at least provides a useful alternative. Summary of the Invention

[0006] In view of the above drawbacks of the prior art, a device for aseptic small volume sampling of liquids is disclosed, comprising:

[0007] A flow control system (FCS), the FCS including a pump; and

[0008] a pipe manifold,

[0009] wherein the pipe manifold includes a first air point, a sample inlet for fluidly coupling the pipe manifold to a cell culture vessel, a second air point, and a sample outlet for fluidly coupling the pipe manifold to a collection point, the pipe manifold being operably coupled to the FCS in use, the FCS including a plurality of valves operable to selectively open and close the first air point, the second air point, the sample inlet, and the sample outlet,

[0010] wherein, in use, the FCS operates the pump and the valves in a forward flow configuration to pump a main fluid from the cell culture vessel into the sample inlet such that a predetermined volume of the main fluid is pumped to the sample outlet and pumped out of the pipe manifold to the collection point, and a portion of the main fluid remains in the pipe manifold,

[0011] wherein, in use, the FCS operates the pump and the valves in a reverse flow configuration to return the portion of the main fluid to the cell culture vessel without exposing the portion of the main fluid to gas.

[0012] There is also disclosed a method for aseptic small volume fluid sampling using a pipe manifold, the pipe manifold including a first air point, a sample inlet fluidly coupled to a cell culture vessel, a second air point, and a sample outlet fluidly coupled to a collection point, the method comprising:

[0013] operating in a forward flow configuration by:

[0014] pumping a main fluid from the cell culture vessel into the pipe manifold via the sample inlet and simultaneously driving air out of the pipe manifold via the second air point,

[0015] pumping the main fluid via the sample inlet, via the pipe manifold, and simultaneously driving air out of the pipe manifold via the sample outlet, and

[0016] pumping gas into the pipe manifold via the first air point and simultaneously driving a predetermined volume of fluid from the main fluid out of the pipe manifold via the sample outlet to the collection point while keeping a portion of the main fluid stationary in the pipe manifold, and

[0017] operating in a reverse flow configuration by: pumping sterile air into the pipe manifold via the second air point and simultaneously driving the main fluid out of the pipe manifold via the sample inlet and back to the cell culture vessel.

[0018] Also disclosed is a method for aseptic collection of a liquid by using a pipeline manifold, wherein the pipeline manifold includes a sample inlet, an air point, a second air point, a sample outlet, and valves at the air point and the sample outlet, and the method includes:

[0019] Closing the valve at the sample outlet and opening the valve at the air point, pumping a predetermined volume of fluid into the pipeline manifold via the sample inlet, and simultaneously driving air to be discharged from the pipeline manifold via the air point.

[0020] Closing the valve at the air point and opening the valve at the sample outlet; and

[0021] Driving a predetermined volume of fluid to be discharged from the pipeline manifold via the sample outlet.

[0022] Also disclosed is a pipeline manifold, which includes a first air pipe, a sample pipe, a second air pipe, a sample outlet pipe, and a pump section fluidly connecting these pipes, wherein the pipeline manifold is configured to engage a flow control system (FCS) such that each of the first air pipe, the second air pipe, the sample pipe, and the sample outlet pipe engages a corresponding valve of the FCS and the pump section engages a pump of the FCS.

[0023] Terms such as "air point", "air inlet", "air outlet", "collection point", "sample inlet", "sample point", "sample outlet (or collection point)" respectively refer to the points or positions in the manifold where air, the main fluid, or the sample can enter or leave the manifold. The functions of these features will become clear from the context of use rather than from the specific terms given for the relevant features. For example, the flow control system can operate in a forward flow configuration or mode and a reverse flow configuration or mode, such that the term "air point" refers to such a feature: air can enter the manifold via this feature in the forward flow configuration, and air can leave the manifold via this feature in the reverse flow configuration. Depending on the context, without losing the generality of the functions that the feature can perform, the same feature can be referred to as "air inlet" or "air outlet".

[0024] The phrases "at the sample inlet" and "at the sample outlet" can mean the place where the pipeline manifold enters / leaves the pipeline housing, the place where it starts to extend away from the FCS, or any other position suitable for providing the functions described herein and as described in the context.

[0025] The phrase "towards the sample outlet" means that air or fluid is pumped via a pipe manifold in the direction of the sample outlet. In the case of fluid, when a predetermined volume of fluid (i.e., the sample to be collected) is pumped towards the sample outlet, the fluid will not be all the fluid pumped into the pipe manifold. Instead, the fluid will be pumped into the pipe manifold until the predetermined volume of fluid is downstream of a valve that can be actuated to open and close the sample point (the fluid flowing downstream from the sample point towards the sample outlet) or downstream of a pump of the fluid control system. Thus, by operating the fluid control system in a countercurrent configuration, any fluid upstream of the predetermined volume of fluid can be returned to the sample point.

[0026] The term "gas" can refer to "air" - for example, "air" from the external environment, "sterile air" or any other suitable gas for the current purpose, especially in cases where the gas maintains the sterility of the bulk fluid. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Embodiments of the present invention will now be described by way of non - limiting examples with reference to the accompanying drawings, in which:

[0028] Figure 1 is a top view of a sterile sampling device according to the present teachings.

[0029] Figure 2 is an exploded schematic view of an automated sterile sampling device according to the present teachings.

[0030] Figure 3 is an assembled automated sterile sampling device, wherein the pipe manifold is located on a pipe panel (currently a single - use or disposable pipe panel), and the pipe panel is locked onto a flow control system (FCS) so as to maintain the alignment (i.e., operatively engage) of the pipe manifold with the FCS.

[0031] Figure 4 is a bottom view of a disposable pipe panel without the pipe manifold.

[0032] Figure 5 shows the pipe manifold.

[0033] Figure 6 illustrates a fully assembled automated sterile sampling device with pipe sensors.

[0034] Figure 7 is for controlling an automated sterile sampling procedure performed by a sterile sampling device using Figure 1 an example graphical user interface (GUI).

[0035] Figures 8 to 12Top view of a fully assembled automated aseptic sampling device with a disposable tubing housing hidden, which respectively includes: a first sensor that draws the main fluid from a cell culture container to a position adjacent to the sample inlet; draws the main fluid through the sample inlet and through a tubing manifold; separates a predetermined volume of fluid from the main liquid; drives the predetermined volume of fluid to be discharged from the tubing manifold through the sample outlet, past a second sensor and to a collection point; and returns a portion of the main fluid remaining in the tubing manifold to the cell culture container.

[0036] Figure 13 Top view of a fully assembled automated aseptic sampling device with a disposable tubing panel hidden, in which the main fluid is directly collected from the cell culture container to the collection point.

[0037] Figure 14 Exploded view of a design variant of an automated aseptic sampling device according to this teaching.

[0038] Figure 15 Is Figure 14 Assembly diagram of the device.

[0039] Figure 16 Is Figure 14 Bottom view of the tubing housing of the device.

[0040] Figure 17 Is Figure 14 Top view of the tubing manifold of the device.

[0041] Figure 18 Shows the device Figure 14 with a sensor attached.

[0042] Figure 19 Is of the device Figure 14 with a disposable tubing panel hidden. Detailed Description

[0043] Disclosed is a device or system for automated aseptic small volume fluid sampling. The device enables an aseptic small volume of fluid (e.g., cell culture medium) to be sampled with substantially no chance of contamination and substantially no fluid loss or waste.

[0044] Figure 1 A device 100 for aseptic small volume sampling of a liquid is shown in. As used herein, a small volume is generally less than 10 ml, such as 20 μl to 1 ml. The device 100 generally includes a flow control system (FCS) 102 and a tubing manifold 104.

[0045] The FCS102 controls the flow of fluid through the pipe manifold 104. In this regard, the FCS102 can be implemented in any suitable configuration to perform the function of driving fluid through the pipe manifold 104 according to the control method described herein.

[0046] Reference Figure 2 , in some embodiments, the FCS200 interacts with the pipe housing 202 to maintain the position of the pipe manifold 204 during use. The FCS200, the pipe housing 202, and the pipe manifold 204 together form an automated aseptic sampling device 206 for sampling small volumes ( Figure 2 the DAAS (Device for Automated Aseptic Sampling) shown in the exploded view of). The pipe housing 202 can be disposable or otherwise single-use, either alone or together with the pipe manifold 204. In this way, the FCS200 can be reused while the components that come into direct contact with the fluid being sampled are disposable. The pipe housing 202 and the pipe manifold 204 can be provided in a single-use kit, which is replaced each time the DAAS206 is used with a fresh cell culture. The single-use kit can be injection-molded from polystyrene (PS), polycarbonate (PC), and / or polypropylene (PP) or any other suitable material. If a transparent finish is required, PS or PC can be used. The kit (pipe housing and pipe manifold) can be produced for use with a specific cell culture (e.g., a cell culture designed to be sized for a specific medium or cell size) to enable pumping of the cell culture without damaging the cells. For packaging, the kit can also be packaged with a cell culture container. For example, the pipe housing and the pipe manifold can be sterilized by using gamma or beta irradiation or ethylene oxide gas (ETO) sterilization. The kit can also include sterile connectors and a cell culture container to enable the kit to be quickly connected to the FCS before sampling.

[0047] On the other hand, the FCS200 is a reusable capital device. The FCS200 is robust and capable of withstanding repeated use. The body 208 of the FCS200 can be made of a sterilizable material (e.g., a material that can be safely placed in an autoclave, such as steel, such as 304 or 316 grade stainless steel), or can be cleaned with 70% ethanol after stripping all its electrical components.

[0048] To ensure that the FCS200 operates accurately when controlling the flow of fluid through the pipe manifold 204, the pipe manifold 204 should be precisely positioned on the FCS200. This can be achieved by providing a groove in the FCS200 corresponding to the shape of the pipe manifold 204, and the pipe manifold 204 is held in place by friction fit, clips, or any other suitable fixing means (e.g., a means capable of removing the pipe manifold 204). However, in Figure 2 In the illustrated embodiment, a kit including the pipe manifold 204 and the pipe housing 202 is used. The accurate positioning and placement of the single-use kit (202, 204) on the FCS200 is achieved by a two-part alignment device, with one part arranged on the FCS200 and the other part arranged on the pipe housing 202 (or on the pipe manifold 204 if the pipe housing 202 is not provided), and the two parts together align the pipe manifold 204 with the FCS200. Currently, alignment is achieved using alignment pins (one of which is labeled 210) arranged on the FCS200. Similarly, alignment can be achieved by positive grooves, notches, protrusions, or other means designed on the FCS200, where the pipe housing 202 has corresponding holes, protrusions, or notches, thus achieving a tight alignment with low tolerances.

[0049] If the two-part system fixes the FCS200 to the pipe manifold 204, then no further fastening system is required. However, in this embodiment, the FCS200 includes a fastening system. The fastening system includes a turn lock (one of which is labeled 212) passing through a corresponding cavity (one of which is labeled 214). If the lock 212 and the cavity 214 are appropriately sized, the alignment device may not be required because the lock 212 and the cavity 214 will align the pipe manifold 204 with the FCS200 and fix it thereto. The fastening mechanism can be configured to allow the pipe manifold 204 to be installed on the FCS 200 unidirectionally (i.e., correctly) only - for example, if the pipe housing 202 is placed on the FCS200 in the wrong way, the cavity 214 may not align with the lock 212.

[0050] Any part of the FCS200 attached to or integrated with the FCS200 can be formed of a robust material to facilitate reuse. This material can be the same as the material forming the FCS200 or, if required, another material.

[0051] The conduit housing 202 is configured to indicate its correct orientation on the FCS 200. Currently, this configuration includes a chamfered edge 215 that indicates the front of the conduit housing 202. This chamfered edge 215 ensures the correct positioning of the pump of the FCS 200 within the corresponding pump cavity 216 of the conduit housing 202. Currently, the pump is a peristaltic pump that includes a roller head 217 fitted into the pump roller head cavity 216. This also ensures the correct positioning of the pump section of the conduit manifold 204 between the conduit housing 202 and the roller head 217 of the pump, applying sufficient pressure to the conduit manifold 204 to enable pumping and driving of fluid through. By ensuring that the conduit manifold 204 and the FCS 200 are aligned in only a single orientation, it is also ensured that the pump drives the fluid in the correct direction and opens and closes the appropriate valves (if any) to facilitate the control of air or gas and fluid.

[0052] The conduit manifold 204 includes a plurality of air points, sample inlets, and sample outlets, through which air or gas can enter and leave the conduit manifold 204. The control of air or gas and sample fluid passing through the conduit manifold 204 is managed by a plurality of valves. These valves are pinch valves, and the heads of the pinch valves are labeled 218. The pinch valves can be part of a device (i.e., the FCS 200) and should be made to ensure robustness to withstand repeated use. The pinch valve heads 218 can be replaceable - for example, after multiple uses - so that the pinch valves can be maintained in the event of wear of the heads 218 after repeated compression cycles. For the pinch valve heads 218, lighter and more robust materials such as polycarbonate (PC), polypropylene (PP), polytetrafluoroethylene (PTFE), and polyamide (NYLON) can be used. These materials, which are manufactured by injection molding, also ensure that they are cost-effective and easy to replace. These plastic materials also work well during the sterilization process using an autoclave or the cleaning process using 70% ethanol.

[0053] Once the conduit housing 202 and the conduit manifold 204 are positioned on the FCS 200 as Figure 3 shown, an alignment mechanism and / or a fixing mechanism can be used to fix the components in their operating configuration (i.e., the components are in an aligned state so that the FCS 200 can be operated to pump fluid through the conduit manifold 204). The turn lock 212 is designed with internal grooves to rotate a predetermined angle of rotation - for example, 90 degrees - specifically to rotate in one of the clockwise or counterclockwise directions for locking and to rotate in the opposite direction for unlocking. Thus, the lock can only be operated in a manner consistent with the correct use of the device. The alignment mechanism and / or the fixing mechanism can ensure the prevention of lateral movement and vertical movement during the use of the device (i.e., during aseptic sampling).

[0054] Refer to Figure 3, the DAAS 300 is configured to attach multiple sensors. In some embodiments, the DAAS 300 itself includes multiple sensors. The DAAS 300 includes a sensor attachment mechanism (currently more than one magnet 302) and an alignment mechanism (currently alignment holes 304). The sensor or sensor block (not shown) (a sensor block is multiple sensors in a single housing) includes corresponding attachment and / or alignment mechanisms to facilitate attachment to the DAAS 300. In the case where the (multiple) sensors or (multiple) sensor blocks are not integrated into the DAAS 300, they can be configured for detachable attachment (e.g., via magnetic attraction to the DAAS 300) and include alignment pins for receiving in the alignment holes 304 to align the (multiple) sensors or (multiple) sensor blocks on the DAAS 300. The DAAS 300 also includes vents (e.g., vent holes 306) for discharging heat generated during use (e.g., from a pump or from a valve). Other heat dissipation systems, such as radiators and fans, can be used.

[0055] The pipe manifold 104 has a first air point 106, a second air point 108, a sample inlet 110, and a sample outlet 112. As Figure 2 shown in the embodiment, the lock 308 is inserted via the alignment aperture 310 to hold the pipe housing 312 in place, thereby fixing the pipe manifold 314 in its operating configuration.

[0056] Figure 4 An embodiment of the pipe manifold 400 is shown. The pipe manifold currently includes a first air point 402, a second air point 404, a sample inlet 406, and a sample outlet 408.

[0057] The components 402, 404, 406, and 408 can be connected in any suitable manner to facilitate the operation of the device (e.g., device 100). In this embodiment, the pipe manifold 400 also includes connectors for connecting the components 402, 404, 406, and 408 together, e.g., Y-shaped connectors 410, 412. As Figure 4 shown, the first air point 402 is connected to the sample inlet 406 via the Y-shaped connector 410, and the second air point 404 is connected to the sample outlet 408. Between the first air point 402 and the sample inlet 406 and the second air point 404 and the sample outlet 408 is a pump section 414. The pump section 414 is aligned with a pump (e.g., the rotating head of a peristaltic pump), so that the pump can drive fluid through the pipe manifold 400.

[0058] The Y-shaped connectors 410, 412 and other components (e.g., air points, inlets and outlets) of the manifold 400 can be made of polycarbonate or polypropylene, which have good compatibility with γ, β radiation or ETO sterilization. Colored polycarbonate or polypropylene can be selected to minimize the discoloration effect caused by γ radiation. The end points 416 can be fitted with connectors to facilitate connection to a cell culture container from which a sample can be extracted. The end points 418 can be similarly fitted with connectors to facilitate connection to a monitoring system for collecting and analyzing samples from the culture container. For example, each connector can be a barb fitting with a Luer connector. The culture container and the monitoring system respectively have corresponding connectors that can be respectively fitted on the two ends 416 and 418, such as Luer connectors. The end point 418 can also be left as an open tube, enabling an operator to collect a sample into a collection container or tube.

[0059] In addition, the entire manifold 400 can have a direct inlet pipeline 406 leading into the cell culture container, a direct output pipeline 408 leading into a monitoring system consumable kit (e.g., it can be a pipe manifold kit directly mounted on the monitoring system), and the air ends 402 and 404 can be connected to a gas filter housing with a filter or any other necessary equipment. For a single cell culture or cell manufacturing process, this entire set of devices can be a disposable single-use device. The entire kit can be packaged and sterilized (e.g., via γ or β radiation or ETO sterilization) as a single unit.

[0060] If necessary, connectors can also be provided for the air points 402, 404. In each case, the connectors can be made of any suitable material (such as polycarbonate or polypropylene) to achieve sterilization via γ or β radiation. Then, a gas filter housing with a corresponding Luer fitting and a filter (e.g., a hydrophobic filter sheet with a pore size of 0.2 μm) can be fitted on the two ends 406 and 408. The gas filter can be pre-sterilized before being attached to the sterilized pipe manifold or packaged together with the pipe manifold for sterilization via γ or β sterilization. This enables sterile air to be drawn in during an automated aseptic sampling process and drives the gas in the tube out.

[0061] The pipe manifold 400 can be directly attached to the FCS (e.g., using a pipe clamp or other mechanisms described herein). The pipe manifold 400 can alternatively be attached by Figure 5is connected to the FCS by the illustrated conduit housing 500. The conduit housing 500 includes a lumen or recess 502. The shape of the cavity 502 is complementary to the outer shape of a portion of the conduit manifold (e.g., is the negative of the outer shape), such that the conduit manifold is positioned within the conduit housing 500 with little to no movement. The fit between the conduit manifold and the conduit housing 500 has a low tolerance fit (i.e., little to no play), or in the illustrated embodiment has a friction fit such that the conduit manifold is slightly compressed to maintain its position within the cavity 502. With respect to the friction fit, the radius of curvature of the inner surface of the cavity 502 is slightly less than the outer radius of the conduit manifold 504 such that the conduit manifold can be snapped into the cavity 502 by the friction fit and ensure that it does not become disengaged from the conduit housing 500. Kits of different sizes can be produced to fit common FCSs or to fit different sized FCSs as needed. For example, the kit can include a conduit housing with an inner radius of 1 / 8” to 1 / 16” and a conduit manifold with an outer radius of 0.25” to 0.188”, where the sizes are selected to match and maintain the friction fit.

[0062] The conduit housing 500 also includes a recess or groove 504 for receiving valves, where the valves are operated to control fluid and gas flow through the conduit manifold (e.g., pinch valve 218) when the conduit housing 500 is aligned on the FCS. The conduit housing 500 also includes corresponding or complementary alignment mechanisms for cooperating with the alignment mechanisms of the FCS to align the FCS and the conduit housing 500. Currently, the alignment mechanism of the conduit housing includes alignment holes 506 for each alignment pin 210.

[0063] Conduit sensors or sensor blocks 600 and 608 (each sensor block including more than one conduit sensor (e.g., sensors for measuring conduit parameters such as flow rate, whether the conduit contains or does not contain fluid, fluid temperature, fluid pressure, etc.)) can be attached to the FCS 602, as Figure 6 illustrated. Attachment can be made by any suitable means, such as alignment pins (not shown) attracted to one or more magnets 302 disposed in the alignment holes 304. Accordingly, the alignment pins are magnetic or are magnets themselves to facilitate attraction to the magnets 304. In other embodiments, the alignment pins are not magnetic. In this way, the alignment pins are separated from the one or more magnets such that the alignment pins ensure that each sensor block is properly aligned with the FCS, where the magnets maintain the engagement between each sensor block and the FCS. In still other embodiments, each sensor block can have a magnet attracted to the FCS, as well as other alignment and attachment configurations that will be apparent to those skilled in the art in view of this teaching.

[0064] The sample inlet 406 is attached to the sensor 604 of the sensor block 600 to sense the presence of fluid in the sample inlet 406 of the conduit manifold 400 - e.g., for fluid entering the conduit manifold 400 via the sample inlet 406. The sample outlet 408 is attached to the sensor 606 of the sensor module 608 to sense the presence of fluid in the sample outlet 408 - e.g., for fluid exiting the conduit manifold 400 via the sample outlet 408. The sensors can also be configured for different conduit sizes.

[0065] A conduit is selected to cooperate with the peristaltic pump (as used herein, the term "cooperate" means that components are shaped, sized, and / or configured to engage with each other to facilitate operation of the device), and valves are enabled to control fluid and gas / air flow through the conduit manifold (along with the pump). The conduit manifold can be formed from any suitable material such as puriflex, c - flex, and ultra - c tubing materials. When the conduit housing and the conduit manifold are fixed to the FCS, the conduit manifold can be fully transparent or transparent in the visible area to assist the sensors 604, 606 in identifying the fluid in the conduit manifold. Materials with a minimal amount of color change due to sterilization (e.g., gamma or beta sterilization) can be similarly used to minimize disruption to sensor operation.

[0066] Further referring Figure 1 ,the sample inlet 110 can be pre - connected to the cell culture vessel 116 prior to sterilization, or can be separately connected after sterilization via a sterile connector on one of the cell culture vessel 116 and the sample inlet 110 or sterile connectors on both (two - part sterile connector 113). The sample outlet 112 can be similarly pre - connected to the collection point 118 prior to sterilization, or can be separately connected after sterilization with sterile connectors 115 pre - attached at both ends. The pinch valve heads 120, 122, 124, 126 are respectively connected to the pinch valves 128, 130, 132, 134 and are operably coupled to the sample inlet 110, the first air point 106, the sample outlet 112, and the second air (including gas) point 108. By positioning each valve in this way, the FCS can direct the cell culture fluid and air flow and generate sample bubbles.

[0067] Information from the sensors described in reference Figure 6 is used by the controller to control the liquid flow. As a result, some sections of the conduit length can vary without the need to reprogram the controller. Assume the sensors are adjacent to the conduit housing or the FCS (or are otherwise at a fixed / known distance from the conduit housing or the FCS): Figure 1(At positions A and B) indicates that the length of the tubing from the first sensor to the cell culture vessel 116 can vary without reprogramming the controller, while the length of the tubing from the second sensor to the collection vessel 118 requires updating if it is changed. Changing the tubing length or minimizing the tubing length reduces the exposure of cell culture fluid not taken from the sample to the external environment of the cell culture vessel.

[0068] During use, valves 120, 122, 124, 126 can be opened under a closed condition, where valve heads 128, 130, 132, 134 compress the tubing manifold 104 against recesses or grooves 504 on the housing 500, thereby sealing the inner cavity of the tubing manifold 104. This prevents air / gas and fluid from flowing into or out of the tubing manifold 104. The collection point 118 can be a series of bags, container manifolds, or other suitable sample collection devices, or a measurement or inspection system directly connected to the device 100 thereon, or open to the atmosphere, enabling the user to collect the effluent fluid using a tube or petri dish.

[0069] Figure 7 An example GUI 700 for controlling the DAAS (device 100) is shown. In addition to programming the FCS for subsequent automated operations, the GUI 700 is also capable of an emergency stop and reset of the sampling process. The FCS can use any suitable mechanism and protocol to accept commands. Currently, the FCS accepts commands via serial communication, making the device compatible as an integrated sampling unit within an automated solution. In the example, the controller of the sampling device is written in the C / C++ programming language, deployed via an Arduino microcontroller, and the GUI 700 is written in Python and deployed on a laptop. The Python interface communicates with the Arduino microcontroller via USB serial communication. The GUI 700 is also capable of manual control of the pinch valves and peristaltic pumps. It enables the user to change parameters such as tubing length, sample volume, and pump speed. If these parameters are not expected to change for a particular application, the default values of these parameters can be set in a configuration file.

[0070] Figures 8 to 12 The figure shows a sterile sampling process using the device 100 according to the present teachings. Once the device, sample container, and sample collection point are in an operating configuration, the process can begin. Initially, as Figure 8As shown, FCS 102 opens valves 124 and 122 (the pinch valves are for illustrative purposes only, and other valves may be used where appropriate, whether those other valves are part of the FCS, the conduit housing, or the conduit manifold). Concurrent with or shortly after opening valves 122, 124, FCS operates peristaltic pump 114 to rotate in a clockwise direction while keeping valves 120, 126 closed. The bulk fluid in cell culture vessel 116 is drawn into conduit manifold 104 and reaches the first conduit sensor 604 of DAAS 100, while concurrently driving air out of the second air point 108. Fluid sensor 604 detects the bulk fluid in the conduit at inlet 110, and FCS 102 records the time it takes for the bulk fluid to travel from cell culture vessel 116 to the first sensor 604.

[0071] The bulk fluid travels through sample inlet 110 of conduit manifold 104 into DAAS 100. When it reaches one end of the first Y-shaped connector 800, FCS 102 determines, based on the program in FCS 102, that the bulk liquid has traveled a predetermined length to reach the transition point from sample inlet 110 to sample outlet 112 (as shown by arrow A - i.e., a predetermined volume of fluid can now be pumped to the location of the sample outlet, where as the predetermined volume of fluid gradually exits the sample outlet, air gradually occupies the conduit manifold), and closes the pinch valve 122 at the second air point 108 while opening the pinch valve 126 at the sample outlet 112, as Figure 9 shown. Pump 114 continues to rotate in a clockwise direction to pump the bulk fluid into the pump section 414 of conduit manifold 104 while driving air out of the sample outlet 112.

[0072] After the bulk fluid has traveled a predetermined distance (along arrow B - the predetermined distance can be calculated based on the flow rate and the dimensions of conduit manifold 104 in the operating configuration, and the flow rate is determined by the full and partial revolutions of the head of pump 114 after fluid is detected at sample inlet 110), FCS 102 determines that the corresponding predetermined sample volume has been reached. In this sense, the predetermined distance is interchangeable with the predetermined volume of fluid pumped past a point where the predetermined volume cannot return to the sample container 116. Thus, the predetermined volume has been pumped into a location in manifold 104 where the predetermined volume can be driven out of sample outlet 112 by sterile air (or gas) entering conduit manifold 104 via the first air point 106. Moreover, all of the bulk fluid except the predetermined volume can be displaced by air returning to cell culture vessel 116 via sample point 110 - for example, only the predetermined volume has moved past the first Y-shaped connector 800, as Figure 10As shown. This also involves closing the pinch valve 124 and opening the pinch valve 120. The pump 114 continues to rotate in the clockwise direction, such that the fluid from the sample pinch valve 124 is separated from the main fluid (indicated by line C) to serve as a predetermined sample volume or better known as a sample bubble (B), and the fluid is facilitated to be discharged from the DAAS 100 via the sample outlet 112 by pumping sterile air from the first air point 106. This occurs when a portion of the main fluid (C) remains stationary at the sample inlet 110 and is sealed by the sample inlet valve 124. The sealing valve 124 helps prevent this portion of the main liquid (C) from contacting the sample outlet 112 or the collection point 118, which helps maintain its sterility.

[0073] Once the sample bubble (B) reaches the second sensor 606 adjacent to the outlet of the DAAS 100 along the sample outlet 112, the FCS 102 determines the predetermined pipe length that the sample bubble must travel to reach the collection point 118. The pump 114 continues to rotate in the clockwise direction to completely drive the sample bubble B out of the pipe manifold 104 and reach the collection point 118.

[0074] Therefore, the FCS 102 drives the sample bubble to the collection point 118. By detecting when the last fluid in the sample bubble (B) passes the sensor 606 - which indicates that the entire sample (B) is now downstream of the valves 126 and the sensor 606 (i.e., towards the collection point 118), it can be determined that all of the sample bubble has been dispensed into the collection point 118. The program in the FCS 102 takes into account the pipe length at the sample outlet 112 and thus operates the pump 114 until all of the sample has been delivered. Therefore, the FCS 102 determines that the sample bubble (B) has completely reached the collection point 118, as Figure 12 shown.

[0075] Then, the FCS 102 activates the countercurrent configuration. Under the countercurrent configuration, the sterile portion of the main fluid (C) returns to the cell culture container 116. This is accomplished by closing the valves 120, 126 at the sample outlet 112 and the first air point 106 and opening the valves 122, 124 at the sample inlet 110 and the second air point 108, as Figure 12As shown. After that, the peristaltic pump 114 is rotated in a countercurrent configuration - i.e., in a counterclockwise manner. This event causes the sterile portion of the bulk fluid (C) to be pumped from the sample inlet 110 to the cell culture vessel 116, while sterile air is drawn into the tubing manifold 104 via the second air point 108. This prevents any fluid or contaminated air from the collection point from being pumped back into the tubing manifold 104, thereby potentially contaminating the bulk fluid. When the bulk fluid passes through the first sensor, the FCS 102 returns this portion of the bulk fluid (C) completely to the cell culture vessel 116 using the time recorded for the bulk fluid to travel from the cell culture vessel 116 to the first sensor 604. Once the FCS 102 determines that there is no longer fluid in the tubing manifold based on the recorded time, it stops the pump 114 and closes all pinch valves 120, 122, 124, 126, and returns to Figure 1 the original configuration in

[0076] In another configuration, the DAAS 100 has been configured to perform an acquisition process of collecting the entire bulk fluid from the cell culture vessel 116 to the collection point 118, as Figure 13 shown. In this case, valves 124 and 126 are opened and valves 120 and 122 are closed, and the pump 114 is operated until the fluid from the sample vessel 116 has passed the sensor 606, and is kept operating until the sensor 606 no longer detects fluid, plus a margin for pumping the fluid through the entire length of the sample outlet 112. This is done only if the collection point 118 already has a separate sterile and sealed tubing manifold (not shown) with a sterile connector for connecting a sterile collection bag, or the collection bag has been pre-connected to the tubing manifold 104. The tube portion of the sampling bag previously used for sampling onto the tubing manifold should be permanently sealed after each sampling process to prevent any contaminants from the sampling bag from potentially contaminating the tubing manifold. If the sampling collection point 118 is exposed to air, another clamp or valve is required between this sample collection point 118 and the point leading to the collection bag and the second sensor. This ensures that the sample collection point 118 can always be in a sealed state after each collection of sample bubbles, while closing the valve 126 at the sample outlet to prevent any contamination of the collection bag or the tubing leading from the second sensor to the collection bag. This can also be done when the collection point 118 is already connected to a measurement system, where the measurement system can ensure that the collected bulk fluid remains sterile and can classify the collected bulk fluid in a sterile manner.

[0077] Open valves 124, 126 while keeping valves 120, 122 closed. Then, rotate pump 114 in the clockwise forward direction to pump the bulk fluid from the cell culture vessel 116 to the collection point 118. The second sensor 606 determines when the bulk fluid leaves the housing, and the FCS 102 determines the predetermined tubing length it must travel to reach the collection collection bag at 118.

[0078] If there are problems with the DAAS 100 or / and the sampling function or / and the collection function, the emergency stop button can be pressed. When the emergency button is pressed, the device closes all pinch valves and stops the peristaltic pump. All other functions are disabled until the system is reset.

[0079] When the reset button is pressed during an emergency stop 702, the cell culture in the cell culture inlet tubing section 110 returns to the cell culture vessel 116. Open the cell culture inlet pinch valve 124 and the second air point pinch valve 122, and the peristaltic pump 114 is set to rotate counterclockwise. If the fluid has not reached the first fluid sensor 604, record the time from the start of the routine until the emergency stop is activated. The pump 114 rotates counterclockwise for the recorded time. If the fluid has passed the first fluid sensor 604, use the fluid sensor 604 to detect the return of the cell culture.

[0080] Then, flush the sample in the cell sample outlet tubing section 112. Close the cell culture inlet pinch valve 124 and valve 122. Then, open valve 120 and the cell sample outlet pinch valve 126, and the peristaltic pump 114 is set to rotate clockwise for a set period of time to completely empty the tubing section. Once the FCS 102 has determined that all fluid has been removed from the tubing manifold 104 (e.g., pumping has reached a predetermined period of time), close all pinch valves and stop the peristaltic pump.

[0081] Figure 14Is an exploded view of the miniaturized DAAS1400. The device 1400 is at least half the length smaller than the DAAS100. The automated aseptic sampling maintains the same configuration and operation as the device 100. The device 1400 still potentially consists of a single-use kit, where the single-use kit includes a tubing housing 1402 and a tubing manifold 1404 that are coupled to the FCS1406 in use. The pump 1408 has been relocated to one end of the DAAS1400, while the pinch valves 1410, 1412, 1414, 1416 for the sample inlet, sample outlet, first air point, and second air point have been collected and positioned at the other end of the DAAS. The pinch valve heads of the pinch valves (one of which is identified as 1418) have also been redesigned for this configuration. Each head 1418 allows a pair of tubes (the sample inlet and the first air point, or the sample outlet and the second air point) to pass through each pinch valve together, while only one of the tubes of the pinch valve is engaged and disengaged by the pinch valve at their respective positions. Thus, each pinch valve has a first condition and a second condition, where in the first condition, the pinch valve engages the first tube (one of the air point, sample inlet, and sample outlet); and in the second state, the pinch valve engages the second tube (one of the air point, sample inlet, and sample outlet). The pinch valves can be moved in any desired manner - for example, the pinch valves can move vertically up and down in the height direction of the FCS (i.e., towards and away from the tubing housing 1402) between the first condition and the second condition. For example, the pinch valve head 1418 and its corresponding pinch valve 1410 will engage or disengage the sample inlet tube, even though both the sample inlet tube and the first air point tube pass through the same pinch valve 1410. This redesigned the DAAS1400 more compactly while maintaining the basic operation and configuration unchanged.

[0082] The negative alignment grooves 1420 have been designed to position and align the kits (1402, 1404) onto the FCS 1406 and lock them against lateral movement. Movement may occur, for example, due to the rotational movement of the peristaltic pump 1408 against the tube 1404 between the pump and the curved pump chamber 1422 in the tube housing 1402. The holes 1424 in the negative grooves 1420 have been threaded to enable the use of knurled or knob screws to fix and lock the single-use kit onto the FCS 1406 via the holes 1426. The screws are used in part to lock the single-use kit against lateral movement, but mainly to prevent vertical movement from the pinch valves 1410, 1412, 1414, 1416. The position and design of the grooves and screws are intended to achieve a compact design of the DAAS 1400. Additionally, in the case of a miniaturized design, fewer fixing and positioning parts are required, thus reducing the number of parts and steps that the user must attend to when assembling the DAAS 1400. The FCS 1406 still includes vent holes 1428 for the peristaltic pump 1408 and the pinch valves 1410, 1412, 1414, 1416 to keep these components cool during long-term and repetitive operation.

[0083] Figure 15 The fully assembled miniaturized DAAS 1400 in includes a single cable hole 1500 for internal components such as the peristaltic pump and the pinch valves. It also includes alignment threaded holes 1502 for sensor block attachment.

[0084] Figure 16 The tube housing 1402 in includes positive alignment grooves or protrusions 1602 for mating with the negative alignment grooves 1420 to lock the kit onto the FCS 1406 and prevent lateral movement. The pinch grooves 1604 are aligned with the pinch valve heads 1418 to clamp or release the tube 1404 located therebetween. The tube cavity 1606 is the same as that of the device 100.

[0085] Figure 17 The tube manifold 1404 in has substantially the same features as the tube manifold 104 (i.e., substantially enabling the same operation despite the modified specific arrangement). The manifold 1404 includes a sample inlet 1702 and a sample outlet 1704 as well as a first air point 1706 and a second air point 1708. The manifold 1404 also includes Y-shaped connectors 1710, 1712 for two pairs of tubes (1706 and 1702, and 1704 and 1708), and a pump section 1714. If Figure 17The pipe manifold therein is formed in a U shape, where the sample outlet 1704 and the second air point 1708 face the same direction as the sample inlet 1702 and the first air point 1706. Then, compared with the straight embodiment where the sample inlet and the first air point face the opposite direction of the sample outlet and the second air point, it looks similar to the Figure 4 original pipe manifold therein.

[0086] Figure 18 shows the Figure 16 pipe housing 1402 and Figure 17 manifold 1404 of Figure 14 and the first sensor 1800 and the second sensor 1802 of the device 1400 in a fully assembled condition. The sensors 1800, 1802 can be directly adhered to the FCS 1406, or can be incorporated into the sensor block 1804 as shown, and then the sensor block 1804 is attached to the FCS 1406 in a manner consistent with the Figure 6 sensor block 600. The sensor block also includes an attachment mechanism (currently the screw holes 1806), and screws can be inserted through the screw holes into the holes 1502 of the FCS 1406 to attach the sensor block 1804 thereto. The sensor block 1804 includes a cavity 1808 for connecting the cables of the two sensors 1800, 1802.

[0087] Figure 19 The pipe manifold 1404 and the pipe housing 1402 in Figure 1 are connected to the cell culture container 1900 via the sample inlet 1702 and to the collection point 1902 via the sample outlet 1704 in a similar manner as shown. The first sensor 1800 is located on the pipe of the sample inlet 1702, adjacent to the inlet to the pipe manifold 1404 within the kit (i.e., the pipe housing 1402 and the pipe manifold 1404, which can together form a disposable or single-use unit). The second sensor 1802 is located on the pipe of the sample outlet 1704, adjacent to the outlet of the pipe manifold 1404. The operation of the device 1400 (with the pipe housing 1402 and the pipe manifold 1404 mounted on the FCS 1406) is the same as that described with reference to Figures 8 to 13 .

[0088] It should be understood that many further modifications and permutations of various aspects of the described embodiments are possible. Accordingly, the described aspects are intended to cover all such changes, modifications, and variations that fall within the spirit and scope of the appended claims.

[0089] Throughout this specification and the following claims, unless the context requires otherwise, the word "comprise", and variations such as "comprises" and "comprising", will be understood to imply the inclusion of the stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.

[0090] Any prior publication (or information obtained therefrom) or known matter referred to in this specification should not be taken as an admission, acknowledgement or any form of implication that such prior publication (or information obtained therefrom) or known matter forms part of the common general knowledge in the technical field to which this specification pertains.

[0091] 1. A device for aseptic small volume sampling of a liquid, comprising:

[0092] A flow control system (FCS); and

[0093] A tubing manifold,

[0094] wherein the tubing manifold includes an air point 1, a sample inlet fluidly connected to a cell culture vessel, an air point 2, and a sample outlet fluidly connected to a collection point, and each of the air point 1, the sample inlet, the air point 2, and the sample outlet is fluidly connected to the tubing manifold and operably connected to the FCS,

[0095] wherein the FCS is capable of operating in a forward flow configuration to pump a bulk fluid from the cell culture vessel to the sample inlet on the tubing manifold,

[0096] wherein the FCS is capable of operating in a forward flow configuration to pump a predetermined volume of fluid from the bulk fluid from the sample inlet to the sample outlet and discharge it from the tubing manifold to the collection point while keeping the bulk fluid stationary,

[0097] wherein, after pumping out a predetermined volume of fluid from the sample outlet and the tubing manifold, the FCS is capable of operating in a reverse flow configuration to return the bulk fluid remaining in the sample inlet on the tubing manifold to the cell culture vessel,

[0098] wherein the forward and reverse flow configurations and operations maintain the sterility of the bulk fluid in the cell culture vessel,

[0099] wherein the forward and reverse flow configurations and operations prevent the presence of dead volume fluid in any part of the tubing from the cell culture vessel to the tubing manifold to the collection point,

[0100] wherein the forward and reverse flow configurations and operations prevent the formation of air bubbles in the fluid leaving the collection point or in the bulk fluid returned to the cell culture vessel.

[0101] 2. The device according to claim 1, wherein the predetermined volume of fluid is a programmable volume of sample bubbles.

[0102] 3. The device according to claim 2, wherein the programmable volume is consistent during repeated sampling processes.

[0103] 4. The device according to claim 2 or / and 3, wherein the range of the programmable volume is from 20 μl to 1 ml.

[0104] 5. The device according to claim 1, wherein the FCS includes valves located at each of air point 1, the sample inlet, air point 2, and the sample outlet, and a pump.

[0105] 6. The device according to claim 5, wherein the FCS is configured to control the valves and actuate the pump to draw the bulk fluid from the cell culture vessel into the tubing manifold via the sample inlet, and simultaneously drive air out of the tubing manifold via air point 2.

[0106] 7. The device according to claims 5 and 6, wherein the FCS opens the valves at the sample inlet and air point 2, closes the valves at the sample outlet and air point 1, and actuates the pump to rotate in a first direction when drawing the bulk fluid from the cell culture vessel into the tubing manifold via the sample inlet and simultaneously driving air out of the tubing manifold via air point 2.

[0107] 8. The device according to claims 5 and 7, wherein the FCS is configured to control the valves and actuate the pump to draw the bulk fluid from the cell culture vessel via the sample inlet, via the manifold, and simultaneously drive air out of the tubing manifold via the sample outlet.

[0108] 9. The device according to claims 5 and 8, wherein the FCS opens the valves at the sample inlet and sample outlet, closes the valves at air point 1 and air point 2, and actuates the pump to rotate in a first direction when drawing the bulk fluid from the cell culture vessel via the sample inlet, via the tubing manifold, and simultaneously driving air out of the tubing manifold via the sample outlet.

[0109] 10. The device according to claims 5 and 9, wherein the FCS is configured to control the valves and actuate the pump to draw sterile air into the tubing manifold via air point 1, and simultaneously drive a predetermined volume of fluid out of the manifold via the sample outlet to a collection point from the bulk fluid while keeping the bulk fluid stationary.

[0110] 11. The device according to paragraphs 5 and 10, wherein the FCS opens the valves at air point 1 and the sample outlet, closes the valves at the sample inlet and air point 2, and actuates the pump to rotate in a first direction when sterile air is drawn into the pipe manifold via air point 1 and, simultaneously, a predetermined volume of fluid is driven out of the pipe manifold via the sample outlet to a collection point from the bulk fluid while keeping the bulk fluid stationary.

[0111] 12. The device according to paragraphs 5 and 11, wherein the FCS is configured to control the valves and actuate the pump to draw sterile air into the pipe manifold via air point 2 and, simultaneously, drive the bulk fluid out of the manifold via the sample inlet back into the cell culture vessel.

[0112] 13. The device according to paragraphs 5 and 12, wherein the FCS opens the valves at air point 2 and the sample inlet, closes the valves at the sample outlet and air point 1, and actuates the pump to rotate in a second direction opposite to the first direction when sterile air is drawn into the pipe manifold via air point 2 and, simultaneously, the bulk fluid is driven out of the manifold via the sample inlet back into the cell culture vessel.

[0113] 14. The device according to paragraph 5, wherein the FCS is configured to control the valves and actuate the pump to draw the bulk fluid from the cell culture vessel into the pipe manifold via the sample inlet and, simultaneously, drive the bulk fluid out of the manifold via the sample outlet to a collection point.

[0114] 15. The device according to paragraphs 5 and 14, wherein the FCS opens the valves at the sample inlet and the sample outlet, closes the valves at air point 1 and air point 2, and actuates the pump to rotate in a first direction when the bulk fluid is drawn from the cell culture vessel into the pipe manifold via the sample inlet and, simultaneously, the bulk fluid is driven out of the manifold via the sample outlet to a collection point.

[0115] 16. The device according to paragraph 1, wherein the FCS includes a first sensor positioned adjacent to the sample inlet and a second sensor positioned adjacent to the sample outlet, wherein the first sensor and the second sensor are configured to detect the presence of fluid at the sample inlet and the sample outlet, respectively.

[0116] 17. The device according to paragraph 16, wherein the FCS is configured to control the forward flow and reverse flow configurations and operations according to the detection of fluid from the first sensor and the second sensor to ensure volume consistency of a predetermined volume of fluid.

[0117] 18. The device according to paragraph 16, wherein the FCS is configured to control the forward flow and reverse flow configurations and operations according to the detection of fluid from the first sensor and the second sensor to ensure sterility of the bulk fluid.

[0118] 19. The device according to claim 16, wherein the FCS is configured to control the co - flow and counter - flow configurations and operations based on the detection of the fluid from the first sensor and the second sensor to prevent the formation of bubbles in a predetermined volume of the fluid exiting the sample outlet or in the bulk fluid returning to the cell culture vessel.

[0119] 20. The device according to claim 16, wherein the FCS is configured to control the co - flow and counter - flow configurations and operations based on the detection of the fluid from the first sensor and the second sensor to prevent the existence of dead volume in any part of the pipeline from the cell culture vessel to the pipeline manifold to the collection point.

[0120] 21. The device according to claim 16, the first sensor is configured to detect when the bulk fluid enters the pipeline manifold via the sample inlet, and the FCS determines when the bulk fluid having a volume enters the pipeline manifold via the sample inlet.

[0121] 22. The device according to claim 21, when subordinate to 6, the FCS opens the valves at the sample inlet and air point 2, closes the valves at air point 1 and the sample outlet, and actuates the pump to rotate in the first direction, thereby pumping the bulk fluid from the cell culture vessel to the first sensor adjacent to the sample inlet, and simultaneously driving the air out of the pipeline manifold via air point 2.

[0122] 23. The device according to claim 22, wherein the FCS records the amount of time taken to pump the bulk fluid from the cell culture vessel to the first sensor.

[0123] 24. The device according to claim 22, when subordinate to 8, the FCS opens the valves at the sample inlet and the sample outlet, closes the valves at air point 1 and air point 2, and when the FCS determines that the bulk fluid has reached a predetermined transition point based on a predetermined distance from the first sensor to the predetermined transition point from the sample inlet to the sample outlet, actuates the pump to rotate in the first direction.

[0124] 25. The device according to claim 24, when subordinate to 8, the FCS opens the valves at the sample inlet and the sample outlet, closes the valves at air point 1 and air point 2, and actuates the pump to rotate in the first direction to pump the bulk fluid via the sample inlet, via the pipeline manifold, and simultaneously drive the air out of the pipeline manifold via the sample outlet.

[0125] 26. The device according to claim 25, when subordinate to 10, the FCS opens the valves at air point 1 and the sample outlet, closes the valves at air point 2 and the sample inlet, and when the FCS determines that a predetermined volume of fluid has been reached since the first detection of the bulk fluid at the first sensor, actuates the pump to rotate in the first direction.

[0126] 27. The device according to 26, when subordinate to 10, the FCS opens the valves at air point 1 and the sample outlet, closes the valves at air point 2 and the sample inlet, and when sterile air is drawn into the pipe manifold via air point 1, and at the same time a predetermined volume of fluid is driven out of the pipe manifold via the sample outlet from the main fluid while keeping the main fluid stationary, actuates the pump to rotate in the first direction.

[0127] 28. The device according to 26 and 16, the second sensor is configured to detect when a predetermined volume of fluid leaves the pipe manifold via the sample outlet, and the FCS determines when a predetermined volume of fluid leaves the pipe manifold via the sample outlet.

[0128] 29. The device according to 28, when subordinate to 10, the FCS opens the valves at air point 1 and the sample outlet, closes the valves at air point 2 and the sample inlet, and when the second sensor detects a predetermined volume of fluid, actuates the pump to rotate in the first direction, and the FCS determines the predetermined pipe length for the predetermined volume of fluid to travel from the second sensor to the collection point.

[0129] 30. The device according to 29, when subordinate to 10, the FCS opens the valves at air point 1 and the sample outlet, closes the valves at air point 2 and the sample inlet, and when sterile air is drawn into the pipe manifold via air point 1, and at the same time a predetermined volume of fluid is driven from the second sensor to the collection point from the main fluid while keeping the main fluid stationary, actuates the pump to rotate in the first direction.

[0130] 31. The device according to 30, when subordinate to 12, the FCS opens the valves at air point 2 and the sample inlet, closes the valves at air point 1 and the sample outlet, and when the FCS has determined that a predetermined volume of fluid has reached the collection point, actuates the pump to rotate in a second direction opposite to the first direction.

[0131] 32. The device according to 31, when subordinate to 12 and 23, the FCS opens the valves at air point 2 and the sample inlet, closes the valves at air point 1 and the sample outlet, and when sterile air is drawn into the pipe manifold via air point 2, and at the same time drives the main fluid to be discharged from the manifold via the sample inlet back into the cell culture container, actuates the pump to rotate in a second direction opposite to the first direction.

[0132] 33. The device according to 32, when subordinate to 23, the FCS is configured to determine when the main fluid has completely returned to the cell culture container and there is no fluid left in the pipes from the cell culture container to the pipe manifold and to the collection point.

[0133] 34. The device according to 33, wherein the FCS closes the air point 1, air point 2, the sample inlet, and the sample outlet, and stops the pump when it is determined that there is no longer fluid in the pipeline from the cell culture container to the pipeline manifold to the collection point.

[0134] 35. The device according to 1, wherein the collection point is connected to a series of sample bags or a container manifold.

[0135] 36. The device according to 35, wherein the countercurrent configuration includes returning the main fluid that has not been contacted by the collection point or the sample bag or container manifold.

[0136] 37. The device according to 36, wherein the countercurrent configuration includes returning the main fluid that has been separated from the sample bubbles to the cell culture container.

[0137] 38. The device according to any one of 1, 2, and 37, wherein the sample bubbles are generated from the main cell culture fluid and separated from the main cell culture fluid.

[0138] 39. The device according to 38, wherein the FCS is configured to seal (e.g., by closing the valve at the sample inlet / sample point) the main cell culture fluid from being exposed to contamination at the collection point.

[0139] 40. The device according to 38, wherein once the sample bubbles are generated, the FCS pumps the sample bubbles from the pipeline manifold to the collection point via the sample outlet.

[0140] 41. The device according to 1, wherein the collection point is exposed to the atmosphere.

[0141] 42. The device according to 1, wherein the collection point is flexibly adapted to the measurement system.

[0142] 43. The device according to 1, wherein the pipeline manifold is carried in a disposable housing that can be attached to the FCS.

[0143] 44. The device according to 43, wherein the housing includes alignment grooves or pins, and the FCS includes corresponding mating holes or notches for the grooves or pins, such that the housing can be positioned on the FCS and locked to prevent lateral movement, wherein the pipeline manifold in the disposable housing is then operably coupled to the peristaltic pump and pinch valves of the FCS.

[0144] 45. The device according to 44, wherein the FCS includes threaded holes, and the housing includes corresponding mating non-threaded holes, such that the housing can be positioned on the FCS and locked by screws or turn locks to prevent lateral movement and vertical movement.

[0145] 46. The device according to claim 43, wherein the pipe manifold is located between the housing and the pinch valve or peristaltic pump of the FCS.

[0146] 47. The device according to claim 46, wherein the housing includes a groove that aligns with the pinch valve when assembled to the FCS, for enabling clamping and releasing of the pipe manifold.

[0147] 48. The device according to claim 47, wherein the housing includes a curved wall that aligns with the peristaltic pump when assembled to the FCS, for enabling pumping of the fluid in the pipe manifold.

[0148] 49. A method for aseptic small - volume sampling of a liquid by using a pipe manifold, wherein the pipe manifold includes an air point 1, a sample inlet fluidly connected to a cell culture container, an air point 2, and a sample outlet fluidly connected to a collection point, and each of the air point 1, the sample inlet, the air point 2, and the sample outlet is fluidly connected to the pipe manifold. The method includes:

[0149] Pumping a main fluid from the cell culture container into the pipe manifold via the sample inlet, and simultaneously driving air out of the pipe manifold via the air point 2,

[0150] Pumping the main fluid via the sample inlet and through the pipe manifold, and simultaneously driving air out of the pipe manifold via the sample outlet,

[0151] Pumping sterile air into the pipe manifold via the air point 1, and simultaneously driving a predetermined volume of fluid out of the pipe manifold via the sample outlet to the collection point from the main fluid while keeping the main fluid stationary, and

[0152] Pumping sterile air into the pipe manifold via the air point 2, and simultaneously driving the main fluid out of the pipe manifold via the sample inlet back to the cell culture container.

[0153] 50. The method according to claim 49, wherein pumping the main fluid from the cell culture container into the pipe manifold via the sample inlet and simultaneously driving air out of the pipe manifold via the air point 2 includes: closing the air point 1 and the sample outlet, and opening the air point 2 and the sample inlet.

[0154] 51. The method according to claim 49, wherein pumping the main fluid via the sample inlet and through the pipe manifold, and simultaneously driving air out of the pipe manifold via the sample outlet includes: closing the air point 1 and the air point 2, and opening the sample inlet and the sample outlet.

[0155] 52. The method according to 49, wherein sterile air is pumped into the pipeline manifold via air point 1 and at the same time a predetermined volume of fluid is driven out of the pipeline manifold via the sample outlet to the collection point from the main body fluid while keeping the main body fluid stationary, including: closing air point 2 and the sample inlet, and opening air point 1 and the sample outlet.

[0156] 53. The method according to 49, wherein sterile air is pumped into the pipeline manifold via air point 2 and at the same time the main body fluid is driven to be discharged from the pipeline manifold via the sample inlet back to the cell culture container, including: closing air point 1 and the sample outlet, and opening air point 2 and the sample inlet.

[0157] 54. A method for collecting a main body fluid using a pipeline manifold, wherein the pipeline manifold includes air point 1, a sample inlet fluidly connected to a cell culture container, air point 2, and a sample outlet fluidly connected to a collection point, and each of air point 1, the sample inlet, air point 2, and the sample outlet is fluidly connected to the pipeline manifold, the method including:

[0158] Pumping a main body volume from the cell culture container into the pipeline manifold via the sample inlet and at the same time driving the main body volume out of the pipeline manifold via the sample outlet to the collection point.

[0159] 55. The method according to 54, wherein pumping a main body volume from the cell culture container into the pipeline manifold via the sample inlet and at the same time driving the main body volume out of the pipeline manifold via the sample outlet to the collection point includes: closing air point 1 and air point 2, and opening the sample inlet and the sample outlet.

[0160] 56. A pipeline manifold, the pipeline manifold includes an air point 1 pipe, a sample inlet pipe fluidly connected to a cell culture container, an air point 2 pipe, a sample outlet pipe fluidly connected to a collection point, and a pump section, wherein the pipeline manifold is configured to engage with an FCS such that each pipe engages with a corresponding valve of the FCS and the pump section engages with the pump of the FCS.

[0161] 57. A pipeline manifold, the pipeline manifold includes an air point 1 pipe, a sample inlet pipe fluidly connected to a sterile connector, an air point 2 pipe, a sample outlet pipe fluidly connected to a sterile connector, and a pump section, wherein the pipeline manifold is configured to engage with an FCS such that each pipe engages with a corresponding valve of the FCS and the pump section engages with the pump of the FCS.

[0162] 58. A device for sterile small-volume sampling of a liquid, including:

[0163] A flow control system, the flow control system includes a pump; and

[0164] A pipeline manifold

[0165] Wherein, the pipe manifold includes a first air point, a sample point, a second air point, and a sample outlet, and each of the first air point, the sample point, the second air point, and the sample outlet is fluidly connected to the pipe manifold and operably connected to a flow control system.

[0166] Wherein, the flow control system (FCS) is capable of operating in a forward flow configuration to pump fluid from the sample point towards the sample outlet.

[0167] Wherein, after pumping a predetermined volume of fluid, the flow control system is capable of operating to pump air from the first air point towards the sample outlet to drive the predetermined volume of fluid out of the manifold via the sample outlet and prevent additional fluid from the sample point from being pumped towards the sample outlet.

[0168] Wherein, after pumping the predetermined volume of fluid out of the manifold, the flow control system is capable of operating in a reverse flow configuration to return the fluid remaining in the pipe manifold to the sample point, and

[0169] Wherein, the forward flow configuration and the reverse flow configuration substantially prevent the formation of air bubbles in the fluid leaving the sample outlet or the sample point.

[0170] 59. The device according to claim 58, wherein the fluid is a programmable volume of sample bubbles.

[0171] 60. The device according to claim 59, wherein the programmable volume is consistent during the repeated sampling process.

[0172] 61. The device according to claim 58 or 59, wherein the range of the programmable volume is from 20 ul to 1 ml.

[0173] 62. The device according to claim 58, wherein the FCS includes valves at each of the first air point, the sample point, the second air point, and the sample outlet, and a pump, and the FCS is configured to control the valves and actuate the pump to draw fluid into the pipe manifold via the sample point and simultaneously drive air out of the pipe manifold via the second air point.

[0174] 63. The device according to claim 62, wherein the FCS closes the valves at each of the first air point and the sample outlet, and actuates the pump to rotate in a first direction when drawing fluid into the pipe manifold via the sample point and simultaneously driving air out of the pipe manifold via the second air point.

[0175] 64. The device according to claim 63, wherein the FCS is configured to control the valves and actuate the pump to draw fluid into the pipe manifold via the sample point and drive a predetermined volume of fluid towards the sample outlet.

[0176] 65. The device according to 64, wherein the FCS closes the valves at each of the first air point and the second air point, and actuates the pump to rotate in a first direction when fluid is drawn into the pipeline manifold via the sample point and a predetermined volume of fluid is driven towards the sample outlet.

[0177] 66. The device according to 65, wherein the FCS is configured to control the valves and actuate the pump to draw air into the pipeline manifold via the first air point and simultaneously drive the fluid to be discharged from the pipeline manifold via the sample outlet.

[0178] 67. The device according to 66, wherein the FCS closes the valves at each of the sample point and the second air point, and actuates the pump to rotate in a first direction when air is drawn into the pipeline manifold via the first air point and simultaneously a predetermined volume of fluid is driven to be discharged from the pipeline manifold via the sample outlet.

[0179] 68. The device according to 67, wherein the FCS is configured to control the valves and actuate the pump to draw air into the pipeline manifold via the second air point and simultaneously drive the remaining fluid in the pipeline manifold to be discharged from the manifold via the sample point.

[0180] 69. The device according to 68, wherein the FCS closes the first air point and the sample outlet, and actuates the pump to rotate in a second direction opposite to the first direction when air is drawn into the pipeline manifold via the second air point and simultaneously the remaining fluid in the pipeline manifold is driven to be discharged from the manifold via the sample point.

[0181] 70. The device according to 69, wherein the FCS includes a first sensor positioned adjacent to the sample point and a second sensor positioned adjacent to the sample outlet, wherein the first sensor and the second sensor are configured to detect the presence of fluid at the sample point and the sample outlet respectively.

[0182] 71. The device according to 70, wherein the FCS is configured to control the forward flow configuration and the reverse flow configuration according to the measurements from the first sensor and the second sensor to prevent the formation of air bubbles at the sample point or the sample outlet.

[0183] 72. The device according to 70, wherein the FCS uses the first sensor and the flow rate corresponding to the speed of the pump to determine when the fluid enters the pipeline manifold, and calculates that a specific volume of fluid not less than the predetermined volume has entered the pipeline manifold from the sample point.

[0184] 73. The device according to 72, wherein the FCS opens the first air point and the sample outlet, closes the sample point and the second air point, and actuates the pump to rotate in a first direction when the first sensor measures that a specific volume of fluid has entered the pipeline manifold.

[0185] 74. The device according to 70, wherein the second sensor is configured to determine when the fluid leaves the pipe manifold, and the FCS determines when a predetermined volume of the fluid leaves the pipe manifold.

[0186] 75. The device according to 74, wherein the FCS closes the first air point and the sample outlet, opens the sample point and the second air point, and actuates the pump to rotate in the second direction when the second sensor measures that a predetermined volume of the fluid has left the pipe manifold.

[0187] 76. The device according to 75, wherein the FCS is configured to determine when there is no fluid in the pipe manifold based on the measurements from the first sensor and the second sensor.

[0188] 77. The device according to 76, wherein the FCS closes the first air point, the second air point, the sample point and the sample outlet, and stops the pump when it determines that there is no fluid in the pipe manifold.

[0189] 78. The device according to 58, wherein the sample outlet is connected to a series of sample bags or a container manifold.

[0190] 79. The device according to 78, wherein the countercurrent configuration includes returning the fluid that has not been contacted by the sample bags or the container manifold or the sample outlet to the sample point.

[0191] 80. The device according to 78 or 79, wherein the countercurrent configuration includes returning the fluid that has been separated from a predetermined volume of the fluid to the sample point.

[0192] 81. The device according to 58, wherein a predetermined volume of the fluid is separated from the main cell culture medium.

[0193] 82. The device according to 81, wherein the flow control system is configured to seal the main cell culture medium to prevent it from being exposed to contamination at the sample outlet. This may involve closing the sample inlet valve. This may also involve separating the sample bubble (i.e., the sample to be delivered to the sample outlet) from the main medium. After the sample bubble is delivered out of the sample outlet, the sample outlet is closed, and a sterile gas (e.g., sterile air from the second air port) can be used to drive the main medium back to the culture container.

[0194] 83. The device according to 59, wherein once a predetermined volume of the fluid forms a sample bubble, the flow control system pumps the predetermined volume of the fluid to the sample outlet.

[0195] 84. The device according to 58, wherein the sample outlet is exposed to the atmosphere.

[0196] 85. The device according to claim 58, wherein the sample outlet is flexibly adapted to different measurement systems.

[0197] 86. The device according to claim 58, wherein the tubing manifold is carried in a disposable housing attachable to an FCS.

[0198] 87. The device according to claim 86, wherein the housing includes an aperture sized to enable the tubing manifold to be positioned on and locked in place on the FCS, operably coupled to a peristaltic pump and pinch valve of the FCS.

[0199] 88. The device according to claim 87, wherein the tubing manifold is positioned between the housing and the pinch valve or peristaltic pump.

[0200] 89. The device according to claim 87, wherein the housing includes a groove that aligns with the pinch valve when assembled to the flow control system to enable clamping and releasing of the tubing manifold.

[0201] 90. A method for aseptic small volume sampling of a liquid using a tubing manifold, wherein the tubing manifold includes a first air point, a sample point, a second air point, and a sample outlet, each of the first air point, sample point, second air point, and sample outlet being fluidly coupled to the tubing manifold, the method comprising:

[0202] pumping a specific volume of fluid into the tubing manifold via the sample point and simultaneously driving air out of the tubing manifold via the second air point,

[0203] pumping air into the tubing manifold via the first air point and simultaneously driving a predetermined volume of fluid not exceeding the specific volume out of the tubing manifold via the sample outlet, and

[0204] pumping air into the tubing manifold via the air outlet and simultaneously driving the remaining fluid in the tubing manifold out of the tubing manifold via the sample inlet.

[0205] 91. The method according to claim 90, wherein pumping a specific volume of fluid into the tubing manifold via the sample point and simultaneously driving air out of the tubing manifold via the second air point includes: closing the first air point and the sample outlet.

[0206] 92. The method according to claim 90, wherein pumping air into the tubing manifold via the first air point and simultaneously driving a predetermined volume of fluid out of the tubing manifold via the sample outlet includes: closing the second air point and the sample point.

[0207] 93. The method according to claim 90, wherein pumping air into the tubing manifold via the second air point and simultaneously driving the remaining fluid in the tubing manifold out of the tubing manifold via the sample point includes: closing the first air point and the sample outlet.

[0208] 94. A method for aseptic collection of a liquid by using a pipe manifold, wherein the pipe manifold includes a sample point, an air point, a sample outlet, and valves at the air point and the sample outlet, and each of the sample point, the air point, the sample outlet, and the valves at the air point and the sample outlet is fluidly connected to the pipe manifold. The method includes:

[0209] Closing the valve at the sample outlet and opening the valve at the air point, pumping a predetermined volume of fluid into the pipe manifold via the sample inlet, and simultaneously driving air out of the pipe manifold via the air point,

[0210] Closing the valve at the air point and opening the valve at the sample outlet; and

[0211] Driving a predetermined volume of fluid out of the pipe manifold via the sample outlet.

[0212] 95. A pipe manifold, which includes a first air pipe, a sample pipe, a second air pipe, a sample outlet pipe, and a pump section, wherein the pipe manifold is configured to engage a flow control system (FCS) such that each of the first air pipe, the second air pipe, the sample pipe, and the sample outlet pipe engages a corresponding valve of the FCS and the pump section engages a pump of the FCS.

Claims

1. An apparatus for aseptic small - volume sampling of a liquid, comprising: Flow control system (FCS), the FCS including a pump; and a piping manifold, wherein the piping manifold includes a first air point, a sample inlet for fluidly coupling the piping manifold to a cell culture vessel, a second air point, and a sample outlet for fluidly coupling the piping manifold to a collection point, the piping manifold being operatively coupled to the FCS in use, the FCS including a plurality of valves operable to selectively open and close the first air point, the second air point, the sample inlet, and the sample outlet, wherein, in use, the FCS operates the pump and the valves in a forward flow configuration to pump a bulk fluid from the cell culture vessel into the sample inlet such that a predetermined volume of the bulk fluid is pumped to the sample outlet and out of the piping manifold to the collection point, and a portion of the bulk fluid remains in the piping manifold, wherein, in use, the FCS operates the pump and the valves in a reverse flow configuration to return the portion of the bulk fluid to the cell culture vessel without exposing the portion of the bulk fluid to gas.

2. The apparatus according to claim 1, wherein, The FCS includes valves at each of the first air point, the sample inlet, the second air point, and the sample outlet.

3. The apparatus according to claim 2, wherein, In the forward flow configuration, the FCS operates the pump and the valves to pump the bulk fluid from the cell culture vessel into the piping manifold via the sample inlet and simultaneously drive gas out of the piping manifold via the second air point.

4. The apparatus according to claim 2 or 3, wherein, The FCS opens the valves at the sample inlet and the second air point, closes the valves at the sample outlet and the first air point, and actuates the pump to rotate in a first direction when pumping the bulk fluid from the cell culture vessel into the piping manifold via the sample inlet.

5. The apparatus according to any one of claims 2 to 4, wherein, The FCS is configured to open the valves at the sample inlet and the sample outlet, close the valves at the first air point and the second air point, and actuate the pump to rotate in a first direction when withdrawing the bulk fluid from the cell culture vessel via the sample inlet and via the piping manifold.

6. The apparatus according to claim 4, wherein, The FCS opens the valves at the sample inlet and the sample outlet, closes the valves at the first air point and the second air point, and actuates the pump to rotate in the first direction when driving the predetermined volume of the bulk fluid out of the manifold to the collection point via the sample outlet.

7. The apparatus according to claim 6, wherein, The FCS is configured to control the valve at the sample inlet to separate the bulk fluid pumped into the manifold into the predetermined volume of fluid and the portion of the bulk fluid, operate the valves and actuate the pump to draw gas into the piping manifold via the first air point and simultaneously drive the predetermined volume of fluid out of the manifold to the collection point via the sample outlet while keeping the portion of the bulk fluid stationary.

8. The apparatus according to claim 7, wherein, The FCS opens the valves at the sample inlet and the second air point, closes the valves at the first air point and the sample outlet, and actuates the pump to rotate in the second direction when driving a portion of the body fluid back into the cell culture vessel.

9. The apparatus according to claim 8, wherein, In the countercurrent configuration, the FCS controls the valves and actuates the pump to draw gas into the pipe manifold via the second air point in a second direction opposite to the first direction, and simultaneously drives a portion of the body fluid to be discharged from the manifold back into the cell culture vessel via the sample inlet.

10. The apparatus according to claim 1, wherein, The FCS includes a first sensor positioned adjacent to the sample inlet and a second sensor positioned adjacent to the sample outlet, wherein the first sensor and the second sensor are configured to detect the presence of fluid at the sample inlet and the sample outlet, respectively.

11. The apparatus according to claim 10, wherein, The FCS is configured to control the forward flow configuration and the countercurrent configuration according to the detection of fluid by the first sensor and the second sensor.

12. The apparatus according to claim 10 or 11, wherein, The FCS further includes a timer and is configured to control the forward flow configuration and the countercurrent configuration according to the detection of fluid from the first sensor and the second sensor and a signal from the timer.

13. The apparatus according to any one of claims 1 to 12, wherein, The pipe manifold includes two Y-shaped connectors. The first Y-shaped connector of the Y-shaped connectors includes the first air point and the sample inlet, and the second Y-shaped connector of the Y-shaped connectors includes the second air point and the sample outlet. The pipe manifold further includes a pump section that engages with the pump in use such that the pump can drive the body fluid into and through the pipe manifold.

14. The apparatus according to any one of claims 1 to 13, further comprising a pipe housing for holding the pipe manifold to the FCS.

15. The apparatus according to claim 12, wherein, The FCS records the amount of time it takes to draw the body fluid from the cell culture vessel to the first sensor.

16. A method for aseptic small - volume fluid sampling using a pipe manifold, the pipe manifold including a first air point, a sample inlet fluidly coupled to a cell culture container, a second air point, and a sample outlet fluidly coupled to a collection point, the method comprising: Operate in the forward flow configuration by: Pumping the body fluid from the cell culture vessel into the pipe manifold via the sample inlet, and simultaneously driving air out of the pipe manifold via the second air point. Pumping the body fluid via the sample inlet and through the pipe manifold, and concurrently driving air out of the pipe manifold via the sample outlet. Pumping gas into the pipe manifold via the first air point, and simultaneously driving a predetermined volume of fluid from the body fluid out of the pipe manifold via the sample outlet to the collection point while keeping a portion of the body fluid stationary in the pipe manifold. Operate in the countercurrent configuration by: pumping sterile air into the pipe manifold via the second air point, and simultaneously driving the body fluid out of the pipe manifold via the sample inlet and back into the cell culture vessel.

17. The method according to claim 16, further comprising sensing the presence of fluid in the pipe manifold at the sample inlet and the sample outlet, and controlling the pumping based on the presence of fluid in the pipe manifold at the sample inlet and the sample outlet.

18. The method according to claim 17, wherein The FCS is configured to control the forward flow configuration and the countercurrent configuration according to the presence of fluid in the pipe manifold at the sample inlet and the sample outlet to prevent the formation of air bubbles at the sample point or the sample outlet.

19. A method for aseptically collecting a liquid by using a pipe manifold, wherein The pipeline manifold includes a sample inlet, an air point, a second air point, a sample outlet, and valves at the air point and the sample outlet, and the method includes: Closing the valve at the sample outlet and opening the valve at the air point, pumping a predetermined volume of fluid into the pipeline manifold via the sample inlet, and simultaneously driving air out of the pipeline manifold via the air point, Closing the valve at the air point and opening the valve at the sample outlet; and Driving the predetermined volume of fluid out of the pipeline manifold via the sample outlet.

20. A pipe manifold comprising a first air pipe, a sample pipe, a second air pipe, a sample outlet pipe, and a pump section fluidly connecting these pipes, wherein The pipeline manifold is configured to engage a flow control system (FCS) such that each of the first air pipe, the second air pipe, the sample pipe, and the sample outlet pipe engages a corresponding valve of the FCS and the pump section engages the pump of the FCS.

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