Bioprocessing equipment

Abstract

The bioprocessing device includes a bioprocessing microfluidic device, a storage tank, a buffer tank for temporary fluid storage, and two fluid communication systems.

Description

Technical Field

[0001] This invention relates to apparatus and methods for processing biological objects, particularly biological cells. Background Technology

[0002] The processing of biological objects plays a vital role in the current development of biotechnology. A complex series of operations, including amplification, concentration, purification, gene editing, gene delivery, RNA delivery, protein delivery, differentiation, dedifferentiation, harvesting, cell sorting, and purification, are common, resulting in compounds that can be directly used for diagnostic or therapeutic purposes or as raw materials for other bioprocessing.

[0003] To achieve high throughput with minimal material usage in these bioprocessing processes, microfluidic devices or microfluidic chips are typically used. In bioprocessing, microfluidic devices require the addition of various reactants or nutrients; cells must be transported, stored, harvested, and so on. These operations require fluid connectivity, which can become extremely complex as the number of microfluidic devices and bioprocessing variants increases.

[0004] U.S. Patent 8,257,964 discloses a microwell cell culture apparatus comprising wells connected to port stations via a multiplexing system. The port stations are intermediate items that provide cells or cell culture medium to each well. The multiplexing system allows for controlled connections between each port station and each well.

[0005] Furthermore, bioprocessing has been developed as a method for producing third- and fourth-generation drugs, including Advanced Therapeutic Drugs (ATMPs). Typically, bioprocessing is optimized within microfluidic systems through screening. Indeed, due to the inherent complexity of living cells, screening is a more feasible approach to optimization than deterministic methods based solely on models.

[0006] Then, scaling from screening batch size to industrial manufacturing batch size becomes particularly complex. In particular, transferring the optimal protocol determined in the screening phase to industrial batch size presents several difficulties: (a) a lack of accuracy in bioprocessing execution during screening can lead to false positives or non-reproducible results; (b) it is generally impossible to scale up the batch size while keeping the protocol parameter values ​​the same. Finally, the high performance of small-scale processes determined in screening is often not reproducible at larger scales, so performance in bioprocessing (e.g., yield, efficiency, product quality, etc.) may anomalously decrease as the batch size increases.

[0007] The object of this invention is to propose a bioprocessing device with a smart fluidic connectivity architecture, in which multiple microfluidic devices can operate simultaneously under specific conditions determined in screening, thereby producing a quantitative amount of compounds of interest. The same bioprocessing device can be used alternately for screening, leveraging the versatility provided by the smart fluidic connectivity architecture to operate different conditions on numerous microfluidic devices. Summary of the Invention

[0008] Therefore, this disclosure relates to a system for processing biological particles, comprising:

[0009] i. At least four bioprocessing microfluidic devices;

[0010] ii. At least three storage devices or at least three ports configured to connect to storage devices;

[0011] iii. at least one buffer tank; and

[0012] iv. At least two fluid-connected systems.

[0013] In addition, the first fluid communication system includes valves and connecting means between the valves, such that each reservoir and a port configured to connect the reservoir can be in fluid communication with each buffer tank; and the second fluid communication system includes valves and connecting means between the valves, such that each bioprocessing microfluidic device can be in fluid communication with each buffer tank.

[0014] In this disclosure, the system uses a storage device to store reactants, suspended biological particles, or, for example, nutrients. The storage device itself may be included in the system, or it may be external to the system but connected to it via a port.

[0015] In this disclosure, the reservoir can be connected and disconnected via a connection port on the flowline. In this case, the connection port is preferably of the type that protects the flowline from contamination of its environment, such as a diaphragm or a swabbable valve. Alternatively, a pipe or delocalized reactant source or product outlet can be used instead of the reservoir. In this disclosure, a flowline is a series of connecting devices, valves, ports, ends, inlets, or outlets that define fluid communication between components of one or more systems.

[0016] In fact, the system needs to be configured to use at least three storage devices: this configuration can be achieved through storage devices, ports configured to connect to storage devices, or a combination thereof. In this disclosure, the term "storage device" includes both storage devices and ports configured to connect to storage devices.

[0017] This system is particularly suitable for processing biological cells, such as white blood cells, T cells, NK cells, hematopoietic stem cells (HSCs), totipotent stem cells, pluripotent stem cells, multipotent stem cells, non-adherent cell lines, and adherent cell lines.

[0018] Various bioprocessing techniques can be performed individually or in combination with this system, such as amplification, concentration, purification, gene editing, gene delivery, RNA delivery, protein delivery, differentiation, dedifferentiation, harvesting, cell sorting, and harvesting and purification.

[0019] The first and second fluid communication systems allow for improved versatility in bioprocessing management. In effect, all reactants can be distributed in a controlled manner within each microfluidic device, reducing the size and dead zone of the connecting and distributing devices. In this disclosure, dead zone refers to the volume of connecting devices that must be filled or flushed with liquid during inflow or outflow from the component (i.e., microfluidic device, buffer tank, or reservoir), the liquid remaining outside the component and lost during transfer.

[0020] In this disclosure, a valve is a device that prevents or allows fluid flow. Valves can be, without limitation, diaphragm valves, wipeable valves (e.g., those disclosed in US6651956), pinch valves, such as those based on elastic tube clamps, pinch valves based on closing microfluidic channels through membrane deformation (e.g., those disclosed in US6929030), other types of membrane-based valves, phase change valves, such as those operated by the liquid contents in a cryotube, mechanical valves (e.g., quarter-turn stopcocks, ball valves), and surface tension-based valves (e.g., in low-pressure applications, an energy barrier is created by simply disconnecting the two parts constituting the flow path due to gas-liquid surface energy). Valves that present a stable closed state are preferred, such as normally closed or bistable valves, to reduce the risk of failure and unwanted flow. Valves compatible with single-use fluid elements (e.g., components in contact with the reaction stream) are generally preferred because having fully single-use fluid elements has been found to reduce the risk of cross-contamination between consecutive batches manufactured using bio-production equipment and to reduce costs. The advantages of miniature valves and conduits that can be integrated into microfluidic devices lie in their reduced dead zone volume and the ability to allow for higher density microfluidic devices and thus higher throughput. Valves also define the boundaries between components throughout the system. In particular, microfluidic devices, reservoirs, and buffer tanks contain fluid inlets and / or outlets. These inlets and / or outlets terminate with valves. Connecting devices are designed between valves to define the liquid distribution from one component to another.

[0021] In one embodiment, the bioprocessing microfluidic device includes at least one chamber for storing and manipulating biological particles, at least one inlet for filling the chamber, and at least one outlet for discharging from the chamber. Due to the inlet and outlet, flow can be applied within the microfluidic chamber without changing its volume. Furthermore, the inlet and outlet can be fluidly communicated with other components of the system via specific connection devices. For example, all reactants can be filled into the microfluidic device through an inlet connected to a first streamline, while all products can be discharged from the microfluidic device through an outlet connected to a second streamline different from the first streamline: reactants and products do not mix in any connection device.

[0022] In the implementation, the chamber receiving the microfluidic device can be sterilized by, for example, autoclaving, gamma ray, or H2O2 vapor.

[0023] Microfluidic devices include at least one port, typically at least two ports. These ports are configured to establish fluid communication between the microfluidic device and a second fluid communication system. Increasing the number of ports allows for the management of complex bioprocesses, but also increases the complexity of connections within the system.

[0024] In certain embodiments, an inlet and an outlet, i.e., two ports, are configured to define a seeding flow. In this disclosure, the seeding flow is a flow that traps biological particles from a liquid flowing into the microfluidic device into a chamber of the microfluidic device, removing biological particles from the liquid flowing out of the microfluidic device during the seeding flow. In one of these embodiments, the trapping of biological particles relies on sedimentation and biological particle precipitation. Various other devices can also be used to facilitate the trapping of biological particles in the microfluidic device, such as flow traps (e.g., pores or micropores smaller than biological particles in the flow) or coatings (e.g., extracellular matrix coatings, antibody coatings, cell adhesion polymer coatings, etc.).

[0025] In another specific embodiment, an inlet and an outlet, i.e., two ports, are configured to define a harvest flow. In this disclosure, the harvest flow is a flow designed to flush the liquid medium of the microfluidic device, causing the contents of the microfluidic device to be discharged for recovery or harvesting. The harvest flow can be gentle enough that the biological particles located in the microfluidic device are not displaced: the discharged liquid may contain the product of interest that is ultimately harvested. The harvest flow can be stronger, thereby displacing and harvesting the biological particles. The main parameters controlling the intensity of the harvest flow are the geometry of the microfluidic device, the viscosity of the liquid, and the flow rate. For example, auxiliary means can be applied to facilitate the harvesting of the product of interest and / or biological particles, such as vibration or ultrasound, chemical treatment, and / or enzymatic treatment.

[0026] In a particular implementation, the inlet and outlet configured to define the inoculation flow are the same as those configured to define the harvest flow. In this implementation, one geometry of the inlet and outlet is used for different flow conditions, such as flow rate, to achieve inoculation or harvesting.

[0027] As disclosed in European patent applications EP18305786 or EP19306568, the two preceding embodiments can be combined in a single microfluidic device to produce a microfluidic device with multiple chambers and fluid channels, allowing for the inoculation and harvesting of these chambers.

[0028] In this implementation, the buffer tank is controlled by a pressure source. The pressure source can generate high pressure that causes the buffer tank to partially or completely displace. The pressure source can also generate low pressure that causes the buffer tank to partially or completely fill. Due to the pressure source, as well as the first and second fluid communication systems, the flow between the system components can be entirely controlled by the pressure source.

[0029] In a particular embodiment, the buffer tank includes at least one chamber, such as a helical tube or coil, a pressure source, and means for controlling the volume of liquid within the chamber. Due to this configuration, the volume within the buffer tank is continuously monitored, allowing for contact with the volume flowing into or out of the microfluidic device or reservoir. A single pressure source and volume monitoring enable control of all volume exchanges within the system. In this embodiment, the average cross-section of the chamber can be 0.1 mm. 2 Up to 9mm 2 This represents a good trade-off between the storage volume of the buffer tank and the hydraulic resistance.

[0030] In a specific implementation, the system for processing biological particles includes at least two buffer tanks, particularly two, three, four, or five buffer tanks.

[0031] In another embodiment, the system for processing biological particles includes at least two buffer tanks, in at least one of which the liquid volume is monitored. With this configuration, a certain volume of first liquid can flow from the first buffer tank to the microfluidic device via a second fluid communication system. Second liquid from the second buffer tank then flows to the same microfluidic device to fill the dead zone of the second fluid communication system, thereby allowing the entire volume of the first liquid to fill the microfluidic device. This is advantageous for small quantities of available reactants or biological particles and avoids wasting valuable biological material on system connectors or inactive components. Alternatively, with this configuration, liquid can flow out of the first buffer tank to collect the contents of the microfluidic device, which simultaneously transfers to the second buffer tank as the volume of the microfluidic device remains substantially constant.

[0032] In the implementation, the buffer tank may be equipped with a bioparticle detector, allowing monitoring of the number of displaced bioparticles, particularly during inoculation of the microfluidic device. Indeed, the number of bioparticles inoculated for bioprocessing is one of the most influential and difficult-to-control parameters. For example, controlling this number allows for determining, for instance, the yield of bioprocessing per particle, which is crucial for screening to select the most promising processes.

[0033] European patent application EP18306872 describes a suitable buffer tank.

[0034] In the implementation scheme, the system for processing bioparticles also includes a waste container, such that each storage container, each buffer container, and each bioprocessing microfluidic device is in fluid communication with the waste container via a first fluid communication system and / or via a second fluid communication system.

[0035] In this implementation, the connection system can be isolated from the waste container via a specific device, such as a one-way check valve. In such an implementation, the connection system to the waste tank can be partially reused and equipped with analytical devices, such as chemical analysis modules, particle suspension analysis modules, or any type of analytical module. Since the flow to the waste container is very frequent during bioprocessing, this implementation allows for periodic analysis of the bioprocessing output fluid, providing extensive information that can be part of the quality control process and used to optimize bioprocessing, for example, by designing corrective actions if certain reactants are above or below certain thresholds. This isolation of the analytical module allows for the use of reusable analytical devices, such as spectrometers, thereby reducing costs.

[0036] In implementations, the connecting device comprises a tube, or a functional equivalent containing channels for fluid flow. Suitable tubes have an inner diameter less than 3 mm, and, depending on the size and configuration of the microfluidic device, are preferably less than 1.6 mm and greater than 0.1 mm. This range of inner diameters represents a good trade-off between the internal volume of the connecting device and hydraulic resistance. Larger inner diameter tubes are preferred for long connections (on the order of meters), and smaller inner diameter tubes are preferred when they are specifically used to connect a reduced number of reservoirs or microfluidic devices. Various types of tubing can be used, generally with medical-grade and relatively inert materials preferred. Silicone, particularly platinum-cured or other USP Class VI compliant silicones, is a suitable choice, although its permeability to gases, especially evaporation through the tube walls, should be considered. PTFE or other fluoropolymers exhibit good properties, particularly in reducing bioparticle adhesion in the connecting device. The tubing's resistance to pressure should be verified, as microfluidic devices may require relatively high infusion pressures. If a material incompatible with pinch valves (e.g., PTFE) is chosen, pinch valves can still be used by employing a small portion of deformable tubing, such as platinum-cured silicone specifically designed for valve-related sections. Regarding the above considerations, the minimized large inner diameter of this section can be used relative to the above considerations, and in particular, the inner diameter of this section can be larger than other types of tubing. This embodiment defines a fixed topology for the connection device between valves, which eliminates the need for complex addressing systems in the first and second fluid communication systems. This embodiment also reduces the risk of contamination of the fluids processed in the connection device by reagents such as atmospheric suspended particles that may be present around the connection device.

[0037] In the implementation, the valve is an end configured to open fluid communication when both ends are in contact and to close fluid communication when one end is not in contact with the other. For example, a suitable end is a combination of a wipeable valve on one side and a connector compatible with a wipeable valve on the other side. The connector connects to an electronically controlled pinch valve or phase change valve—that is, a device that blocks flow as close as possible to the distal end of the connector. In this example, the electronically controlled pinch valve or phase change valve is more complex and has a wider range of integration, so the arrangement of the wipeable valve and connector should minimize the number of connectors and maximize the number of wipeable valves. For example, the wipeable valve would be the end directly associated with the microfluidic device, while the connector coupled to the electronically controlled valve would be the end directly associated with the buffer tank. This approach is particularly advantageous because the connection topology is dynamically established, allowing the elimination of fixed connection devices, such as tubing, thereby reducing the volume of the connection devices and thus reducing the dead zone and / or volume occupied by the connection devices in the system.

[0038] In implementations with dynamic topologies, mechanical actuations of any type of microfluidic device can be used individually or in combination, relative to the buffer tank. This allows for combinations of 1D, 2D, or 3D arrays of microfluidic devices, facilitating high-density integration.

[0039] Systems used for processing biological particles can use either a fixed topology or a dynamic topology.

[0040] In this embodiment, the internal volume of the second connection system is less than 300% of the total volume of all bioprocessing microfluidic devices. Such a volume is desirable to prevent valuable reactants or bioparticles from remaining in the connection devices instead of entering the active components of the system, i.e., the microfluidic devices. In this disclosure, the internal volume of an element is the volume of liquid that can be contained within that element. The internal volume of the second connection system is the sum of the volumes of each connection device within the connection system.

[0041] In the implementation scheme, the number of valves in the first fluid communication system is less than three times the number of reservoirs multiplied by the number of buffer tanks, preferably twice the number of buffer tanks, and more preferably multiplied by the number of buffer tanks.

[0042] In one implementation, the number of valves in the second fluid communication system is less than the total number of ports of all bioprocessing microfluidic devices multiplied by the number of buffer tanks. In a specific implementation, the number of valves in the second fluid communication system is less than the total number of bioprocessing microfluidic devices multiplied by the number of buffer tanks.

[0043] In a typical multiplexing system, each reservoir is connected to each microfluidic device via a valve-controlled connection. Therefore, the required number of valves is the number of microfluidic devices multiplied by the number of containers. Six microfluidic devices and ten reservoirs would require 60 valves. In practice, because they are multiple ports on each microfluidic device, typical multiplexing systems allow each port of each microfluidic device to be individually connected to any reservoir. The required number of valves is then the number of ports per microfluidic device multiplied by the number of containers. Typically, microfluidic devices have at least two ports, and often four.

[0044] The introduction of buffer tanks allows for a reduction in the number of valves, as connections only need to be established between all microfluidic devices and a small number of buffer tanks, as well as between all reservoirs and a small number of buffer tanks.

[0045] In the two previous implementations, reducing the number of valves was desirable because it corresponded to a reduction in complexity, namely a reduction in the number of system components and a reduction in the overall volume of the connecting devices between valves.

[0046] In one implementation, the number of valves in the first and second fluid communication systems is less than the total number of ports of all bioprocessing microfluidic devices multiplied by the number of reservoirs. In a specific implementation, the number of valves in the first and second fluid communication systems is less than the total number of bioprocessing microfluidic devices multiplied by the number of reservoirs.

[0047] In this embodiment, some components of the system for processing bioparticles are enclosed in a pressurized chamber, particularly the microfluidic device and optionally a buffer tank and a second fluid communication system. In this embodiment, a pneumatic clamping force is applied to each microfluidic device present in the system, i.e., a pressure difference is generated between the pressure of the clamping fluid in the system and the pressure of the fluid (applied by the pump and flow) in the microfluidic device. In the case of microfluidic devices including an elastomeric brake base plate and / or an elastomeric cover plate, the clamping force is also advantageous because it reduces the pressure difference between the inside and outside of the microfluidic device during use, thereby reducing deformation of the elastomeric material and limiting changes in the geometry of the microfluidic device. Furthermore, such a pressure difference ensures that no flow escapes from the microfluidic device in the event of a leak, which is particularly advantageous when the fluid contains hazardous or valuable materials. Finally, pneumatic clamping offers significant advantages over mechanical clamping systems, such as rigid plates with bolts, C-clamps, magnets, or shafts and levers, which restrict or impede access to the periphery of the microfluidic device. Conversely, pneumatic clamping provides access to the microfluidic device across its entire periphery, increasing the possibility of establishing fluid communication or optically monitoring the contents of the microfluidic device.

[0048] European patent application EP19306048 describes a suitable pressurization chamber.

[0049] This disclosure also relates to methods for processing biological particles using the system described above, the methods including:

[0050] i. Liquid containing biological particles flows from at least one reservoir into at least one buffer tank via a first fluid communication system; and

[0051] ii. and to allow liquid containing biological particles to flow from at least one buffer tank into at least one bioprocessing microfluidic device through a second fluid communication system.

[0052] This method is particularly suitable for processing biological cells, such as leukocytes, T cells, NK cells, hematopoietic stem cells (HSCs), totipotent stem cells, pluripotent stem cells, multipotent stem cells, non-adherent cell lines, and adherent cell lines. Attached Figure Description

[0053] The features and advantages of the present invention will become apparent from the following description of embodiments of the system and method according to the present disclosure, which is given by way of example only and with reference to the accompanying drawings, wherein:

[0054] Figure 1 It is a schematic architecture of a system for processing biological particles in a fixed topology of a connecting device.

[0055] Figure 2 It is a schematic architecture of a system for processing biological particles, in which part of the connecting devices are in a fixed topology and another part of the connecting devices are in a dynamic topology.

[0056] Figure 3 This is a schematic architecture of a system for processing biological particles, in which the dynamic topology of the connecting devices is achieved by moving microfluidic devices. Detailed Implementation

[0057] Figure 1 A system (1) according to a first embodiment of the present disclosure is shown, designed for processing biological particles. Six microfluidic devices (20) are placed in a chamber (2) of the system (1). Each microfluidic device includes an inlet and an outlet (i.e., two ports), both terminated by a valve (502). Ten reservoirs (40) are placed in the system (1), and each reservoir (40) contains an outlet terminated by a valve (502). Here, the reservoirs (40) are refrigerated in a refrigeration chamber (4). Four buffer tanks (30) are placed in the system (1), and each buffer tank (30) contains an inlet / outlet terminated by a valve (502). Here, the buffer tanks (30) in the chamber (3) are temperature-controlled, typically at the temperature for processing biological cells. Connecting devices (501) in the form of tubular pipes are arranged between the valves (502). By appropriate configuration of opening and closing the valves, each reservoir can be in fluid communication with each buffer tank, and each buffer tank can be in fluid communication with each microfluidic device.

[0058] In this application, a buffer tank is a fluid element in which liquid is introduced, temporarily stored, and then discharged. For example, a buffer tank can be a chamber or a long, narrow tube.

[0059] Here, the first fluid communication system includes valves (502) associated with the reservoirs (40) and buffer tanks (30) and connection devices (501) between these valves (502). Twenty-eight valves (502) are used to connect ten reservoirs (40) to four buffer tanks (30). The second fluid communication system includes valves (502) associated with the microfluidic device (20) and buffer tanks (30) and connection devices (501) between these valves (502). The valves (502) associated with the buffer tanks (30) are part of both the first and second fluid communication systems.

[0060] The microfluidic device (20) is also connected to control modules (22, 23) to control the temperature and dissolved gas concentration in the chamber (2). The water content of the microfluidic device is further controlled by module (24) to measure water loss and ultimately add or remove water from the microfluidic device as needed. When evaporation results in water loss, water vapor is added to the chamber containing the microfluidic device (20).

[0061] As shown in a non-limiting manner, the system (1) includes a waste tank (42) which is in fluid communication with each reservoir (40), each buffer tank (30), and each microfluidic device (20). In the illustrated embodiment, a set of connection devices (501) closest to the reservoir (40), buffer tank (30), and microfluidic device (20) (through an inlet) is used to flow the contents of the reservoir (40) into the microfluidic device (20) through temporary storage in the buffer tank (30), defining a first flow path. A set of connection devices (501) furthest from the reservoir (40), buffer tank (30), and microfluidic device (20) (through an outlet) is used to flow liquid into the waste tank (42), thereby defining a second flow path. With this configuration, liquid treatment of the waste tank (42) does not use the same connection devices (501) as liquid delivery to the microfluidic channel (20).

[0062] Furthermore, in the illustrated embodiment, the connection system includes two separate flow lines connecting the microfluidic device (20) to either a buffer tank (30) or a waste tank (42). With this configuration, liquid can flow from the first buffer tank (30) to collect the contents of the microfluidic device (20), which are simultaneously transferred to a second buffer tank (30) since the volume of the microfluidic device remains substantially constant. Having at least two buffer tanks (30) that can be connected to a single microfluidic device (20) via different flow lines allows for the recollection of liquid from the microfluidic device, particularly when it contains products of interest or biological particles, for transfer to an external container connected via a port or to a storage tank (40).

[0063] exist Figure 1In the example shown, the buffer tank (30) is controlled by a pressure source (311), which is a pressure controller in this case. By depressing the pressure source (311), liquid flows from the reservoir (40) or the microfluidic device (20) into the buffer tank (30). As the pressure of the pressure source (311) increases, liquid is directed from the buffer tank (30) to the microfluidic device (20), the reservoir (40), or the waste tank (42). To avoid bubble formation caused by low pressure, it is preferable to use increased pressure from the pressure source. In a specific case where pressure is applied from the first buffer tank (30) to the microfluidic device (20) to collect the contents of the microfluidic device (20) into the second buffer tank (30), the first buffer tank (30) is pressurized and the second buffer tank (30) is maintained at a sufficiently high pressure to avoid bubble formation.

[0064] According to this implementation of system (1), the controller (10), which has a user interface (11) and a central computer (101), is able to set the flow in the system according to the considered bioprocessing. The controller monitors parameters such as temperature, pressure, humidity, gas concentration in the microfluidic device, water loss of the microfluidic device, time and duration of process steps, and defines the flow between all components of the system according to the flow rate and displacement volume.

[0065] In one variant, the system (1) may consist of a plurality of chambers (2), each chamber (2) containing at least four bioprocessing microfluidic devices; having a plurality of chambers (4), each chamber (4) containing at least three reservoirs (40) or at least three ports configured to connect to the reservoirs; and a plurality of chambers (3), each chamber (3) containing at least one buffer tank (30).

[0066] This variant is typically obtained by adding fluid communication between two subsystems, each subsystem being like... Figure 1 As shown. For example, a storage device (40) can be replaced by fluid communication between two subsystems.

[0067] This variant increases the system's versatility. Bioprocessing microfluidic devices can be stored at different temperatures while using the same storage container. Depending on the stored chemicals, several storage chambers can also be controlled at different temperatures. Several buffer solutions can be used for specific steps in the liquid flow, avoiding cross-contamination. Last but not least, this parallelization variant allows for the on-demand increase in the number of bioprocessing microfluidic devices used under similar conditions.

[0068] Figure 2A system (1) according to a second embodiment of the present disclosure is shown, designed for processing biological particles. Elements similar to those in the first embodiment have the same reference numerals. Six microfluidic devices (20) are placed in a chamber (2) of the system (1). Each microfluidic device includes two ports: an inlet end (505) and an outlet end (505) serving as valves. Ten reservoirs (40) are placed in the system (1), and each reservoir (40) contains an end (505) serving as a valve. Here, the reservoirs (40) are refrigerated in a refrigeration chamber (4). Four buffer tanks (30) are placed in the system (1), and each buffer tank (30) contains an inlet / outlet ending with a valve (502). Here, the buffer tanks (30) in the chamber (3) are temperature-controlled, typically at the temperature for processing biological cells. A tubular connection (501) is arranged between the valves (502). The two ends (505) serving as valves are arranged on tubes within two syringes (506). The first fluid communication system includes a valve (502), a syringe (506), and terminals (505) associated with the buffer tank (30) and a reservoir, and a connection device (501) between these valves / terminals. The second fluid communication system includes a valve (502), a syringe (506), and terminals (505) associated with the buffer tank (30) and a microfluidic device (20), and a connection device (501) between these valves / terminals.

[0069] As shown in a non-limiting manner, a buffer tank (30) and a pressure source (311) are placed on a moving head (510), the displacement of which is controlled by an arm (511). Through appropriate movement of the moving head (510), the tip (505) of a syringe (506) contacts the tip (505) of a reservoir (40), thereby opening the fluid communication of the first fluid communication system. Then, after another movement of the moving head (510), the tip (505) of a syringe (506) contacts the tip (505) of a microfluidic device (20), thereby opening the fluid communication system of the second syringe.

[0070] exist Figure 2 In the example shown, two fluid connections are simultaneously established between the microfluidic device (20) and the two syringes (506). One fluid connection is used to flow liquid from a first buffer tank (30) to the microfluidic device (20), and the second fluid connection is used to flow liquid from the microfluidic device (20) to a second buffer tank (30), thereby maintaining a constant volume in the microfluidic device (20). Liquid removed from the microfluidic device (20) is stored in the buffer tank (30) and can be discarded into a waste container (42) or further used in bioprocessing in another microfluidic device (20) or stored as a final product in a storage container (40).

[0071] In this embodiment, the volume of the connecting device (501) is very strictly limited because the topology of the connecting device (501) is dynamically adjusted according to the displacement requirements of the moving head (510). In particular, the internal volume of the second connecting system is independent of the number of microfluidic devices (20) or the distance between them. Therefore, the volume transferred from the buffer tank (30) to the microfluidic device is almost completely transferred, with no liquid remaining in the dead zone. Furthermore, only 20 valves / pipettes are needed to connect 10 tanks to 4 buffer tanks. 22 valves / terminals are used to connect 6 microfluidic devices, each with two ports and 4 buffer tanks per port. A total of 32 valves / terminals are sufficient to connect 10 reservoirs and 6 microfluidic devices with multiple flows and processes.

[0072] In this embodiment, the chamber (2) is pressurized such that the pressure in the chamber (2) is higher than the pressure in the microfluidic device (20). This overpressure avoids any risk of leakage through the ends (505). When the two ends (505) come into contact, the high or low pressure generated by the pressure source (311) is sufficient to allow liquid to flow through the ends (505).

[0073] Figure 3 A system (1) according to a third embodiment of this disclosure is shown, designed for processing biological particles. Elements similar to those in the first and second embodiments have the same reference numerals. Twelve microfluidic devices (20) are placed in a chamber (2) of the system (1). Each microfluidic device includes two ports: an inlet end (505) and an outlet end (505) serving as valves. Ten reservoirs (40) are placed in the system (1), and each reservoir (40) contains an outlet ending with a valve (502). Here, the reservoirs (40) are refrigerated in a refrigeration chamber (4). Four buffer tanks (30) are placed in the system (1), and each buffer tank (30) contains an inlet / outlet ending with a valve (502). Here, the buffer tanks (30) in the chamber (3) are temperature-controlled, typically at the temperature for processing biological cells. Connecting devices (501) in the form of tubular tubes are arranged between the valves (502). The two ends (505) serving as valves are arranged on tubes within two fixed syringes (506). The first fluid communication system includes valves (502), syringes (506), and terminals (505) associated with the buffer tank (30) and the reservoir, as well as connection means (501) between these valves / terminals. The second fluid communication system includes valves (502), syringes (506), and terminals (505) associated with the buffer tank (30) and the microfluidic device (20), as well as connection means (501) between these valves / terminals.

[0074] As shown in a non-limiting manner, a moving head (510), whose displacement is controlled by an arm (511), can hold and move the microfluidic device (20) at different positions within the chamber (1). Through appropriate movement of the moving head (510), the ends (505) of two syringes (506) contact the two ends (505) of the microfluidic device (20), thereby opening a second fluid communication system, similar to the second embodiment. The syringes (506) may be mounted on mechanical actuators and / or equipped with detectors, such as contact or pressure sensors, allowing for feedback-regulated coupling. The topology of the second fluid communication system can be dynamically adjusted as needed. On the other hand, the first fluid communication system is similar to the first embodiment. This embodiment is particularly relevant when using a large number of microfluidic devices (20), for example, more than 100, while using very few reservoirs (40) in the system (1).

[0075] exist Figure 3 In the example shown, an additional bioprocessing module (7) is provided within the chamber. This additional module can be used for selective processing on a microfluidic device (20) moved by a moving head (510), such as washing, cell sorting (optical, magnetic, or by size exclusion), electroporation, filtration, lysis, microinjection, purification, ion exchange, or any commonly used bioprocessing steps, such as amplification, concentration, purification, gene editing, gene delivery, RNA delivery, protein delivery, differentiation, dedifferentiation, harvesting, cell sorting, and harvesting and purification.

[0076] exist Figure 3 In the example shown, an additional analysis module (8) is set up in the system (1). The microfluidic device (20) can be moved within the analysis module (8) via a moving head (510), and then the microfluidic device (20) itself or the liquid contained therein can be analyzed by microscopy, spectroscopy, mass spectrometry, chemical analysis, rheology, polymerase chain reaction (PCR), reverse transcription polymerase chain reaction (RT-PCR), ELISA, gene sequencing, or any commonly used analytical protocol. Specifically, the microfluidic device (20) can be used as a low-capacity reservoir for transfer to the analysis module (8) for analysis. The microfluidic device (20) can be configured for specific analyses.

Claims

1. A system for processing biological particles, the system comprising: i. At least four bioprocessing microfluidic devices (20); ii. At least three storage devices (40) or at least three ports configured to connect to storage devices; iii At least one buffer tank (30); and iv. At least two fluid-connected systems; The first fluid communication system includes valves (502) and connecting means (501) between valves (502), such that each reservoir (40) or a port configured to connect a reservoir can be in fluid communication with each buffer tank (30); and The second fluid communication system includes valves (502) and connecting devices (501) between valves (502), enabling each bioprocessing microfluidic device (20) to be in fluid communication with each buffer tank (30).

2. The system for processing biological particles according to claim 1, further comprising at least one waste container (42), The second fluid communication system includes valves (502) and connecting devices (501) between the valves (502), enabling each bioprocessing microfluidic device (20) to be in fluid communication with each buffer tank (30) or waste tank (42), and The connection system includes two independent flow lines, a first flow line and a second flow line. The first flow line connects the storage unit (40) to the microfluidic device (20) through the buffer tank (30), and the second flow line connects each microfluidic device (20) and each buffer tank (30) to the waste tank (42).

3. The system for processing biological particles according to claim 1, further comprising a waste container (42) such that each reservoir (40), each buffer container (30) and each bioprocessing microfluidic device (20) is in fluid communication with the waste container (42) via a first fluid communication system and / or via a second fluid communication system.

4. The system for processing biological particles according to claim 1, wherein the connecting device (501) comprises a tube.

5. The system for processing biological particles according to claim 1, wherein the valve is an end (503) configured to open fluid communication when the two ends (503) are in contact, and configured to close fluid communication when one end (503) is not in contact with the other end (503).

6. The system for processing biological particles according to claim 1, wherein the internal volume of the second connection system is less than 300% of the volume of all the bioprocessing microfluidic devices (20).

7. The system for processing biological particles according to claim 1, wherein the number of valves (502) in the first fluid communication system is less than three times the number of reservoirs (40) multiplied by the number of buffer tanks (30).

8. The system for processing biological particles according to claim 1, wherein the number of valves (502) in the second fluid communication system is less than the number of ports of all bioprocessing microfluidic devices (20) multiplied by the number of buffer tanks (30).

9. The system for processing biological particles according to claim 1, wherein the number of valves (502) in the first fluid communication system and the second fluid communication system is less than the number of ports of all bioprocessing microfluidic devices (20) multiplied by the number of storage units (40).

10. The system for processing biological particles according to claim 1, wherein the buffer tank (30) is controlled by a pressure source (311).

11. The system for processing biological particles according to claim 1, wherein the system comprises at least two buffer tanks.

12. The system for processing biological particles according to claim 1, wherein the microfluidic device (20) is enclosed in a pressurized chamber (2).

13. A method for processing biological particles using the system according to any one of the preceding claims, the method comprising: i. The liquid containing biological particles flows from at least one reservoir (40) into at least one buffer tank (30) through a first fluid communication system; and ii. The liquid containing biological particles is fed from at least one buffer tank (30) into at least one bioprocessing microfluidic device (20) through a second fluid communication system.

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