Bioprocessing Equipment

Abstract

The bioprocessing device includes a bioprocessing microfluidic device, a reservoir, a buffer tank for temporarily storing fluids, and two fluid communication systems.

Description

[Technical Field]

[0001] The present invention relates to an apparatus and method for processing biological material, particularly biological cells. [Background technology]

[0002] The processing of biological materials is strongly linked to current biotechnological developments: a complex series of operations such as amplification, enrichment, purification, gene editing, gene delivery, RNA delivery, protein delivery, differentiation, dedifferentiation, recovery, cell sorting, and recovery and purification are common, resulting in compounds that can be used directly for diagnostic or therapeutic purposes or as raw materials for other biological processes.

[0003] To operate with minimal amounts of material and enable high throughput in these biological processes, microfluidic devices or chips are often used. During biological processes, the microfluidic devices need to be supplied with various reactants or nutrients; cells need to be transported, stored, and recovered. These operations require fluid communication and can become very complex with the increasing number of variants of microfluidic devices and biological processes.

[0004] U.S. Patent No. 8,257,964 discloses a microwell cell culture device including wells that communicate with port stations via a multiplexing system. The port stations are intermediate items for supplying cells or cell culture medium to each well. The multiplexing system allows for separately controlled communication between each port station and each well.

[0005] In addition, biological processes are being developed as production methods for third- and fourth-generation drugs, the latter including advanced therapy medicinal products (ATMPs). In many cases, biological processes are optimized in microfluidic systems by screening. Indeed, as a result of the very complexity of biological cells, screening is a more viable means of optimization than model-based deterministic approaches alone.

[0006] Therefore, scaling up from a screening batch size to an industrial manufacturing batch is particularly complex. In particular, transferring a protocol identified as optimal in the screening stage to an industrial batch size is associated with several difficulties: (a) lack of precision in the execution of the biological process during screening can lead to false positives or irreproducible results, and (b) increasing the batch size while maintaining the same values ​​of the protocol parameters is usually not possible. Finally, high-performance small-scale processes identified in screening usually cannot be reproduced at a larger scale, and thus, paradoxically, the performance (e.g., yield, efficacy, product quality...) may decrease in a biological process when the batch size is increased.

[0007] The objective of the present invention is to propose a bioprocessing device in which many microfluidic devices can be simultaneously operated under specific conditions identified in screening to produce a fixed amount of a target compound using an excellent fluid communication structure. Alternatively, the same bioprocessing device can be used in screening, taking advantage of the versatility provided by the excellent fluid communication structure to perform different conditions in many microfluidic devices. Summary of the Invention

[0008] Accordingly, the present disclosure provides a system for processing biological particles, comprising: i. at least four bioprocessing microfluidic devices; ii. at least three reservoirs or ports configured to communicate with the reservoirs; iii. at least one buffer tank; iv. at least two fluid communication systems; Including, Regarding the system.

[0009] Furthermore, the first fluid communication system includes valves and communication means between the valves such that each reservoir or port configured to communicate with the reservoir can be fluidly connected to each buffer tank, and the second fluid communication system includes valves and communication means between the valves such that each bioprocessing microfluidic device can be fluidly connected to each buffer tank.

[0010] In the present disclosure, the system uses a reservoir to store, for example, reactants, biological particles, or nutrients in suspension. The reservoir may be contained within the system itself, or the reservoir may be external to the system and communicated with the system via a port.

[0011] In the present disclosure, reservoirs can be connected and disconnected via communication ports on flow lines. In such cases, the communication ports are preferably of a type that protects the flow lines from contamination from their environment, such as septa or swabable valves. Reservoirs can also be replaced by, for example, pipes or delocalized reactant sources or product outlets. In the present disclosure, a flow line is a collection of communication means, valves, ports, chips, inlets or outlets that define one or several fluid connections between components of the system.

[0012] In practice, the system must be configured to use at least three reservoirs; this configuration can be achieved with reservoirs, ports configured to communicate with reservoirs, or a combination thereof. In this disclosure, the term "reservoir" encompasses both reservoirs and ports configured to communicate with reservoirs.

[0013] The system 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 cells, and adherent cell lines.

[0014] Various biological processes, such as amplification, enrichment, purification, gene editing, gene delivery, RNA delivery, protein delivery, differentiation, dedifferentiation, recovery, cell sorting, and recovery and purification, may be performed alone or combined with this system.

[0015] The first and second fluid communication systems allow for improved versatility in the management of biological processes. Indeed, all reactants may be distributed to each microfluidic device in a controlled manner, reducing the size and dead portion of the communication and distribution means. In the context of this disclosure, dead portion refers to the volume of the communication means that must be filled with or flushed with liquid during flow into or out of a component, i.e., a microfluidic device, buffer tank, or reservoir. The liquid remains outside the component and is lost in translation.

[0016] In the present disclosure, a valve is a means for preventing or allowing flow. Valves can be, but are not limited to, septa or swabable valves (e.g., as disclosed in U.S. Patent No. 6,651,956), pinch valves, such as those based on pinching elastic tubing, or those based on closing a microfluidic channel by membrane deformation (e.g., as disclosed in U.S. Patent No. 6,929,030), other types of membrane-based valves, phase-change valves, such as valves activated by freezing the liquid content in the tubing, mechanical valves (e.g., quarter-turn stoppers, ball valves), or surface tension-based valves (e.g., valves that simply disconnect two sections of a flow path upon application of low pressure, creating an energy barrier due to the surface energy of the air and liquid). To reduce the risk of failure and unwanted flow, valves that exhibit a stable closed state, such as normally closed or bistable valves, are preferred. Valves compatible with single-use fluidic elements (e.g., components in contact with reactant streams) are generally preferred, as having completely disposable fluidic elements has been found to reduce the risk of cross-contamination between successive batches produced in a biological production tool and also reduces costs. Miniaturized valves and valves that can be integrated into microfluidic devices are advantageous because they reduce dead volume, enable high-density microfluidic devices, and allow for higher throughput. Valves also define boundaries between components of the overall system. In particular, microfluidic devices, reservoirs, and buffer tanks contain inlets and / or outlets for fluids. These inlets and / or outlets terminate at valves. Communication means are designed between the valves to define fluid flow from one component to another.

[0017] In one embodiment, a bioprocessing microfluidic device includes at least one chamber in which bioparticles can be stored and manipulated; at least one inlet for loading the chamber and an outlet for discharging the chamber. The inlet and outlet allow flow to be imposed on the microfluidic chamber without changing its volume. Furthermore, the inlet and outlet may be fluidically connected to other components of the system via specific communication means. For example, all reactants may be loaded into the microfluidic device through an inlet connected to a first flow line, and all products may be discharged from the microfluidic device through an outlet connected to a second flow line different from the first flow line; the reactants and products are not mixed via any communication means.

[0018] In one embodiment, the chamber receiving the microfluidic device can be sustained for sterilization, for example, by autoclaving, gamma radiation, or H2O2 vapor.

[0019] A microfluidic device includes at least one port, and typically at least two ports, configured to establish fluid communication between the microfluidic device and a second fluid communication system. Increasing the number of ports allows for more complex biological processes to be managed, but increases the complexity of the communication in the system.

[0020] In certain embodiments, one inlet and one outlet, i.e., two ports, are configured to define a seeding flow. In the present disclosure, the seeding flow is a flow that captures bioparticles in a chamber of the microfluidic device from a liquid flowing into the microfluidic device; during the seeding flow, the liquid flowing from the microfluidic device is devoid of the bioparticles that flowed into the device. In one of these embodiments, bioparticle capture relies on sedimentation, where the bioparticles settle. Various other means, such as flow traps (e.g., openings in the flow, microwells, smaller than the bioparticles) or coatings (e.g., extracellular matrix coatings, antibody coatings, cell adhesion polymer coatings, etc.), can also be used to facilitate bioparticle capture in microfluidic devices.

[0021] In another specific embodiment, one inlet and one outlet, i.e., two ports, are configured to define a recovery flow. In the present disclosure, a recovery flow is a flow intended to flow the liquid medium of the microfluidic device, thereby discharging and recovering (recovering or harvesting) the contents of the microfluidic device. The recovery flow may be gentle enough not to displace bioparticles in the microfluidic device; the discharging liquid may contain the desired product and is ultimately recovered. The recovery flow may be strong enough to displace and recover bioparticles. The main parameters determining the strength of the recovery flow are the geometry of the microfluidic device, the liquid viscosity, and the flow rate. Auxiliary measures, such as vibration or ultrasound, chemical and / or enzymatic treatment, may be applied to assist in the recovery of the desired product and / or bioparticles.

[0022] In certain embodiments, the inlet and outlet configured to define the seeding flow are the same as the inlet and outlet configured to define the recovery flow, in which one inlet and outlet geometry is used with different flow conditions, e.g., flow rate, to achieve seeding or recovery.

[0023] Both of the preceding embodiments may be combined into a single microfluidic device, as disclosed in European Patent Application Nos. 18305786 or 19306568, resulting in a microfluidic device with several chambers and fluid channels that allow seeding and recovering these chambers.

[0024] In one embodiment, the buffer tank is controlled by a pressure source. The pressure source provides a high pressure, which may result in partial or total draining of the buffer tank. The pressure source provides a low pressure, which may result in partial or total filling of the buffer tank. With the pressure source and the first and second fluid communication systems, flows between the components of the system may all be controlled by the pressure source.

[0025] In certain embodiments, the buffer tank comprises at least one chamber, e.g., in the form of a spiral tube or coil; a pressure source; and a means for controlling the volume of liquid in the chamber. With this configuration, the volume in the buffer tank is monitored, providing continuous access to the volume flowing to or from the microfluidic device or reservoir. A single pressure source and volume monitoring allows for control of all volume exchanges in the system. In this embodiment, the chamber is 0.1 mm 2 ~9mm 2 which is a good compromise between the storage content and hydraulic resistance of the buffer tank.

[0026] In certain embodiments, the system for processing biological material comprises at least two buffer tanks, in particular 2, 3, 4 or 5 buffer tanks.

[0027] In another embodiment, a system for processing biological particles includes at least two buffer tanks, the liquid volume of which is monitored in at least one of them. In such a configuration, a volume of a first liquid can be flowed from a first buffer tank through a second fluid communication system to a microfluidic device. A second liquid can then be flowed from the second buffer tank to the same microfluidic device, thereby filling the inactive portion of the second fluid communication system and allowing the entire volume of the first liquid to be loaded into the microfluidic device. This is preferred for reactants or biological materials that are available in low quantities and to avoid wasting valuable biological materials into the communication means or inert components of the system. Alternatively, with such a configuration, the contents of the microfluidic device can be recovered by flowing liquid from the first buffer tank, which is then simultaneously transferred to the second buffer tank, since the volume of the microfluidic device remains essentially constant.

[0028] In one embodiment, the buffer tank may be equipped with a bioparticle detector, which allows monitoring the number of displaced bioparticles, especially during seeding of the microfluidic device. In fact, the number of bioparticles seeded in a bioprocess is one of the most influential and difficult-to-control parameters. Controlling this number allows, for example, determining the yield of the bioprocess per particle, which is important in screening to select the most promising processes.

[0029] A suitable buffer tank is described in European Patent Application No. 18306872.

[0030] In one embodiment, the system for processing biological particles further includes a waste tank such that each reservoir, each buffer tank, and each bioprocessing microfluidic device can be in fluid communication with the waste tank via the first fluid communication system and / or the second fluid communication system.

[0031] In one embodiment, the communication system can be separated from the waste container by specific means, such as a one-way check valve. In such an embodiment, the communication system connected to the waste tank can be partially reused and can include analytical means, such as a chemical analysis module, a particle suspension analysis module, or any type of analytical module. In fact, the flow to the waste container is very frequent over the life of the bioprocess, so such an embodiment can be part of a quality control process, allowing for regular analysis of the output fluid of the bioprocess, providing a wide range of information that can be used to optimize the bioprocess; for example, if some reactants exceed or fall below a certain threshold, corrective measures can be programmed. This separation of the analytical module allows for the use of reusable analytical cells, such as spectroscopic cells, resulting in lower costs.

[0032] In one embodiment, the communication means comprises tubing or a functionally equivalent element containing a channel for liquid flow. Suitable tubing may have an internal diameter of less than 3 mm, with internal diameters of less than 1.6 mm and greater than 0.1 mm being preferred, depending on the size and configuration of the microfluidic device. This range of internal diameters represents a good compromise between the internal volume and hydraulic resistance of the communication means. Larger internal diameter tubing is preferably used for long connections (measured in meters), while smaller internal diameter tubing is preferred when they are specific to the connection of a small number of reservoirs or microfluidic devices. Various types of tubing materials may be used, with medical-grade materials and relatively inert substances generally preferred. Silicone, particularly platinum-cured silicone or other USP Class VI compliant silicones, is a suitable choice, although its permeability to gases should be considered, particularly with regard to associated evaporation through the tubing wall. PTFE or other fluorinated polymers exhibit good performance, particularly for reducing adhesion of biological particles to the communication means. The tubing's resistance to pressure should be verified, as microfluidic devices may require relatively high perfusion pressures. If a material, such as PTFE, is selected that is incompatible with pinched-type tubing valves, pinched-type tubing valves can still be used by using a small section of deformable tubing, such as platinum-cured silicone, for the segment connected to the valve. This section is minimized, and a larger inner diameter can be used for this section, particularly for other types of tubing. This embodiment defines a consistent topology of the communication means between the valves, eliminating the need for complex display systems in the first and second fluid communication systems. This embodiment also reduces the risk of contamination of the fluids operated through the communication means by agents, such as airborne particles, that may be present in the surrounding communication means.

[0033] In one embodiment, the valve is a chip configured to open fluid communication when two chips contact each other and close fluid communication when the chip is not in contact with another chip. A suitable chip is, for example, a combination of a wipeable valve and a compatible connector on one side of the wipeable valve. The connector is coupled to an electronically controlled pinch valve or phase transition valve, i.e., a means for blocking flow as close as possible to the distal end of the connector. In this example, the electronically controlled pinch valve or phase transition valve is more complex and extensive for integration, where the placement of the wipeable valve and connector minimizes the number of connectors and maximizes the number of wipeable valves. For example, the wipeable valve is a chip directly coupled to the microfluidic device, and the connector coupled to the electronically controlled valve is a chip directly coupled to a buffer tank. This embodiment is particularly advantageous because the topology of the communication means is dynamically established, allowing for the elimination of fixed communication means such as tubing, thereby reducing the volume of the communication means and reducing the dead mass and / or volume occupied by the communication means of the system.

[0034] In embodiments with dynamic topologies, all kinds of mechanical actuation of microfluidic devices relative to buffer reservoirs can be used alone or in combination, with combinations that allow for arranging 1D, 2D, or 3D arrays of microfluidic devices being preferred for high density integration.

[0035] A system for processing bioparticles may use a constant topology in one part and a dynamic topology in another part.

[0036] In one embodiment, the internal volume of the second communication system is less than 300% of the volume of the entire bioprocessing microfluidic device. Such a volume is desirable to avoid valuable reactants or biological particles remaining in the active components of the system, i.e., the communication means, instead of proceeding to the microfluidic device. In this disclosure, the internal volume of an element is the volume of liquid that can be contained in this element. The internal volume of the second communication system is the sum of the volumes of each communication means contained in the communication system.

[0037] In one embodiment, the number of valves in the first fluid communication system is less than three times the number of buffer tanks, preferably less than two times the number of buffer tanks, more preferably less than the number of reservoirs multiplied by the number of buffer tanks.

[0038] In one embodiment, the number of valves in the second fluid communication system is less than the number of ports of all bioprocessing microfluidic devices multiplied by the number of buffer tanks. In certain embodiments, the number of valves in the second fluid communication system is less than the number of ports of all bioprocessing microfluidic devices multiplied by the number of buffer tanks.

[0039] In a typical multiplexing system, each reservoir is connected to each microfluidic device via a valve-controlled communication means. Thus, the number of valves required is the number of microfluidic devices multiplied by the number of reservoirs. For six microfluidic devices and ten reservoirs, 60 valves are required. In effect, these are multiple ports on each microfluidic device, so a typical multiplexing system allows each port on each microfluidic device to be individually connected to any reservoir. Thus, the number of valves required is the number of ports on each microfluidic device multiplied by the number of reservoirs. Typically, a microfluidic device has at least two ports, and often four ports.

[0040] The introduction of buffer tanks allows for a reduced number of valves, since it is only necessary to establish communication between all microfluidic devices and a small number of buffer tanks, and between all reservoirs and a small number of buffer tanks.

[0041] In both of the above embodiments, a reduction in the number of valves is desirable as this corresponds to a reduction in complexity, i.e., the number of elements of the system, and a reduction in the overall volume of the communication means between the valves.

[0042] In one embodiment, the number of valves in the first and second fluid communication systems is less than the number of ports of all bioprocessing microfluidic devices multiplied by the number of reservoirs. In certain embodiments, the number of valves in the first and second fluid communication systems is less than the number of bioprocessing microfluidic devices multiplied by the number of reservoirs.

[0043] In one embodiment, some components of the system for processing bioparticles are contained in a pressurized chamber, particularly a 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, resulting from the pressure difference between the pressure of the clamping fluid in the system and the pressure of the fluid in the microfluidic device (imposed by the pump and flow). In the case of microfluidic devices including an elastomeric backplate and / or an elastomeric cover plate, the clamping pressure can also reduce the pressure difference between the inside and outside of the microfluidic device during use, thereby reducing deformation of the elastomeric material and limiting variations in the geometry of the microfluidic device. Furthermore, such a pressure difference ensures that flow can be prevented from exiting the microfluidic device in the event of a leak in the microfluidic device, which is particularly advantageous when the fluid contains hazardous or rare substances. Finally, pneumatic clamping is highly advantageous compared to mechanical clamping systems such as bolts, C-clamps, magnets, or rigid plates including shafts and levers, which limit or prevent access to the periphery of the microfluidic device. In contrast, when using pneumatic clamping, access to the microfluidic device is provided around its entire periphery, increasing the possibility to establish fluid communication or to optically monitor the contents of the microfluidic device.

[0044] A suitable pressurised chamber is described in European Patent Application No. 19306048.

[0045] The present disclosure also provides a method for processing biological particles using the above-described system, comprising: i. flowing a liquid containing bioparticles from at least one reservoir through a first fluid communication system to at least one buffer tank; ii. flowing the liquid containing the biological particles from the at least one buffer tank through a second fluid communication system to the at least one bioprocessing microfluidic device; The present invention relates to a method, comprising:

[0046] The method is suitable for treating biological cells, particularly biological cells such as leukocytes, T cells, NK cells, hematopoietic stem cells (HSCs), totipotent stem cells, pluripotent stem cells, multipotent stem cells, non-adherent cells, and adherent cell lines.

[0047] Features and advantages of the present invention will be apparent from the following description of embodiments of the systems and methods according to the present disclosure, which description is given by way of example only and with reference to the accompanying drawings. [Brief explanation of the drawings]

[0048] [Figure 1] 1 is a schematic structure of a system for processing biological particles with a constant topology configuration of communication means. [Figure 2] 1 is a schematic structure for processing biological particles with one part of a communication means of fixed topology and another part of a communication means of dynamic topology. [Figure 3] The dynamic topology of the communication means is a schematic structure of a system for processing bioparticles achieved by moving a microfluidic device. DETAILED DESCRIPTION OF THE INVENTION

[0049] FIG. 1 shows a system (1) according to a first embodiment of the present disclosure, intended for processing biological particles. Six microfluidic devices (20) are arranged in a chamber (2) of the system (1). Each microfluidic device includes an inlet and an outlet (i.e., two ports), both of which terminate in a valve (502). Ten reservoirs (40) are arranged in the system (1) and include outlets terminating in valves (502). Here, the reservoirs (40) are refrigerated in a refrigerated chamber (4). Four buffer tanks (30) are arranged in the system (1) and include inlet / outlet ports terminating in valves (502). Here, the buffer tanks (30) are temperature-controlled in the chamber (30) at temperatures typically used for processing biological cells. Communication means (501) in the form of tubes are arranged between the valves (502). By appropriate configuration of open and closed valves, each reservoir can be in fluid communication with a respective buffer tank, and each buffer tank can be in fluid communication with a respective microfluidic device.

[0050] In this application, a buffer tank is a fluid element into which a liquid is introduced, temporarily stored, and then discharged. The buffer tank can be, for example, a chamber or an elongated tube.

[0051] Here, the first fluid communication system includes valves (502) coupled to the reservoirs (40) and buffer tanks (30) and communication means (501) between these valves. Twenty-eight valves (502) are used to communicate the ten reservoirs (40) with the four buffer tanks (30). The second fluid communication system includes valves (502) coupled to the microfluidic device (20) and buffer tanks (30) and communication means (501) between these valves (502). The valve (502) coupled to the buffer tanks (30) is part of both the first and second fluid communication systems.

[0052] The microfluidic device (20) is further connected to control modules (22, 23) for temperature and concentration of dissolved gases within the chamber (2). The water content of the microfluidic device is further controlled by a module (24) for measuring water loss in the microfluidic device and adding or removing water accordingly, as needed. If water loss is due to evaporation, water vapor is added to the chamber containing the microfluidic device (20).

[0053] As shown in this non-limiting example, the system (1) includes a waste tank (42) that can be in fluid communication with each reservoir (40), each buffer tank (30), and each microfluidic device (20). In the illustrated embodiment, the set of communication means (501) located closest (via inlets) to the reservoirs (40), buffer tanks (30), and microfluidic devices (20) is used to direct the contents of the reservoirs (40) to the microfluidic devices (20) via temporary storage in the buffer tanks (30), defining a first flow line. The set of communication means (501) located farthest (via outlets) from the reservoirs (40), buffer tanks (30), and microfluidic devices (20) is used to direct the liquid to waste (42), defining a second flow line. In such a configuration, liquids that can be disposed of in waste (42) do not use the same communication means (501) as liquid delivery to the microfluidic channel (20).

[0054] Additionally, in the illustrated embodiment, the communication system includes two independent flow lines connecting the microfluidic device (20) to either the buffer tank (30) or the waste tank (42). In such a configuration, liquid can be flowed from the first buffer tank (30) and the contents of the microfluidic device (20) can be withdrawn, which is simultaneously transferred to the second buffer tank (30) since the contents of the microfluidic device remain essentially constant. Having at least two buffer tanks (30) that can be connected to a single microfluidic device (20) via different flow lines allows liquid to be withdrawn again from the microfluidic device, particularly if it contains a product or bioparticle of interest, for transfer to an outlet container or reservoir (40) connected via a port.

[0055] In the embodiment shown in FIG. 1 , the buffer tank (30) is controlled by a pressure source (311), here a pressure controller. Restriction of the pressure source (311) induces a flow of liquid from the reservoir (40) or the microfluidic device (20) to the buffer tank (30). Increasing the pressure of the pressure source (311) induces a flow of liquid from the buffer tank (30) to the microfluidic device (20), the reservoir (40), or waste (42). Increasing the pressure of the pressure source is preferably used to avoid bubble formation induced by low pressure. In the specific case of imposing a flow from the first buffer tank (30) to the microfluidic device (20) to recover 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 held at a pressure high enough to avoid bubble formation.

[0056] In this embodiment of the system (1), a controller (10) with a user interface (11) and a central computer (101) allows the configuration of flows in the system according to the intended biological process. The controller monitors parameters: temperature, pressure, humidity, gas concentration of the microfluidic device, water loss of the microfluidic device, time and duration of processing steps, and defines flows between all components of the system in terms of flow rates and displaced volumes.

[0057] In a variant, the system (1) may be organized with a plurality of chambers (2), each chamber (2) containing at least four bioprocessing microfluidic devices; a plurality of chambers (4), each chamber containing at least three reservoirs (40) or ports configured to communicate with reservoirs; and a plurality of chambers (3), each chamber (3) containing at least one buffer tank (30).

[0058] This variant is usually obtained by adding fluid communication between two subsystems, each of which is illustrated in Figure 1. For example, one reservoir (40) can be replaced by a fluid communication between both subsystems.

[0059] This variant increases the versatility of the system. Bioprocessing microfluidic devices can be stored at different temperatures while using the same reservoirs. The chambers of some reservoirs can also be controlled at different temperatures depending on the chemicals stored. Some buffers can be used for specific steps of the liquid flow, avoiding cross-contamination. Last but not least, this parallelization variant allows for an increased number of bioprocessing microfluidic devices used under similar conditions, on demand.

[0060] FIG. 2 shows a system (1) according to a second embodiment of the present disclosure, intended for processing biological particles. Similar elements of the first embodiment bear the same references. Six microfluidic devices (20) are arranged in the chamber (2) of the system (1). Each microfluidic device includes two ports: an inlet tip (505) acting as a valve and an outlet tip (505). Ten reservoirs (40) are arranged in the system (1) and include tips (505) acting as valves. Here, the reservoirs (40) are refrigerated in the refrigerated chamber (4). Four buffer tanks (30) are arranged in the system (1) and include inlet / outlet ports terminating in valves (502). Here, the buffer tanks (30) are temperature-controlled in the chamber (3), typically at temperatures for processing biological cells. Between the valves (502) are arranged communication means (501) in the form of tubes. Two chips (505) acting as valves are arranged in tubes between two injectors (506). The first fluid communication system includes the valve (502), the injector (506), the chip (505) connected to the buffer tank (30) and the chip (505) connected to the reservoir, and a communication means (501) between these valves / chips. The second fluid communication system includes the valve (502), the injector (506), the chip (505) connected to the buffer tank (30) and the chip (505) connected to the microfluidic device (20), and a communication means (501) between these valves / chips.

[0061] As illustrated in a non-limiting example, the buffer tank (30) and the pressure source (311) are disposed on a moving head (510), the displacement of which is controlled by an arm (511). By appropriate movement of the moving head (510), the tip (505) of one injector (506) is brought into contact with the tip (505) of the reservoir (40), thereby opening fluid communication in a first fluid communication system. Then, after another movement of the moving head (510), the tip (505) of one injector (506) is brought into contact with the chip (505) of the microfluidic device (20), thereby opening fluid communication in a second fluid communication system.

[0062] In the example shown in Figure 2, two fluidic connections are simultaneously established between the microfluidic device (20) and two injectors (506). One fluidic connection is used to flow liquid from the first buffer tank (30) into the microfluidic device (20), and the second fluidic connection is used to flow liquid from the microfluidic device (20) to the 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 may be discarded to waste (42), further used in a biological process in another microfluidic device (20), or stored in the reservoir (40) as a final product.

[0063] In this embodiment, the volume of the communication means (501) is very strongly limited because the topology of the communication means (501) is dynamically adapted according to the displacement and demands of the moving head (510). In particular, the internal volume of the second communication system is independent of the number of microfluidic devices (20) and the distance between them. Thus, the volume transferred from the buffer tank (30) to the microfluidic devices is almost completely transferred, without leaving any liquid in inactive areas. Furthermore, only 20 valves / chips are used to communicate four buffer tanks and ten reservoirs. Also, 22 valves / chips are used to communicate six microfluidic devices with two ports, each with four buffer tanks. A total of 32 valves / chips is sufficient to communicate ten reservoirs and six microfluidic devices with excellent flow and process versatility.

[0064] In this embodiment, chamber (2) is pressurized so that the pressure in chamber (2) is higher than the pressure in the microfluidic device (20). This excess pressure avoids any risk of leakage through chip (505). When two chips (505) come into contact, the high or low pressure provided by pressure source (311) is sufficient to force liquid through chip (505).

[0065] FIG. 3 shows a system according to a third embodiment of the present disclosure, intended for processing biological particles. Elements similar to those in the first and second embodiments have the same reference numerals. Twelve microfluidic devices (20) are arranged in a chamber (2) of the system (1). Each microfluidic device includes two ports: an inlet tip (505) acting as a valve and an outlet tip (505). Ten reservoirs (40) are arranged in the system (1) and include an outlet terminating in a valve (502). The reservoirs (40) are refrigerated in a refrigerated chamber (4). Four buffer tanks (30) are arranged in the system (1) and include an inlet / outlet terminating in a valve (502). The buffer tanks (30) are temperature-controlled in the chamber (3) at temperatures typically used for processing biological cells. Communication means (501) in the form of tubes are arranged between the valves (502). Two chips (505) acting as valves are arranged in tubes between two fixed injectors (506). The first fluid communication system includes a valve (502) and reservoir connected to a buffer tank (30), an injector (506), and a chip (505), and a communication means (501) between these valves / chips. The second fluid communication system includes a chip (505) connected to the valve (502), the injector (506), and the buffer tank (30), and a chip (505) connected to the microfluidic device (20), and a communication means (501) between these valves / chips.

[0066] As presented in a non-limiting example, a transfer head (510), whose displacement is controlled by an arm (511), can hold and move the microfluidic device (20) at different positions in the chamber (1). By appropriate movement of the transfer head (510), the tips (505) of both injectors (506) are brought into contact with the two tips (505) of the microfluidic device (20), thus opening the fluid communication of the second fluid communication system, as in the second embodiment. The injectors (506) may be mounted on mechanical actuators and / or equipped with detectors, such as contact or pressure sensors, allowing for feedback coupling adjustment. The topology of the second fluid communication system is dynamically adapted as required. Meanwhile, the first fluid communication system is similar to the first embodiment. This embodiment is particularly relevant when a large number of microfluidic devices (20), e.g., more than 100, are used and only a few reservoirs (40) are used in the system (1).

[0067] In the example shown in Figure 3, an additional bioprocessing module (7) is disposed in the chamber, which can be used for selective processes in one microfluidic device (20) moved by the moving head (510), such as washing, cell sorting (optical, magnetic, or molecular sieving), electroporation, filtration, lysis, microinjection, purification, ion exchange, or any conventional bioprocessing step, such as amplification, concentration, purification, gene editing, gene delivery, RNA delivery, protein delivery, differentiation, dedifferentiation, recovery, cell sorting, and recovery and purification.

[0068] In the example shown in Figure 3, an additional analytical module (8) is disposed in the system (1). The microfluidic device (20) may be transferred to the analytical module (8) by the transfer head (510), whereupon the microfluidic device (20) itself or the liquid contained therein may be analyzed by microscopy, spectroscopy, mass spectrometry, chemical analysis, rheology, polymerase chain reaction (PCR), reverse transcription PCR (RT-PCR), ELISA, genetic sequencing, or any conventional analytical protocol. Specifically, the microfluidic device (20) may be used as a small-volume reservoir for transfer to the analytical module (8) and subsequent analysis. The microfluidic device (20) may be configured for a specific analysis.

Claims

1. 1. A system for processing biological particles, comprising: i. at least four bioprocessing microfluidic devices (20); ii. at least three reservoirs (40) or ports configured to communicate with the reservoirs; iii. at least one buffer tank (30); iv. at least two fluid communication systems; Including, the first fluid communication system includes valves (502) and communication means (501) between the valves (502), and each reservoir (40) or a port configured to communicate with the reservoir is in fluid communication with each buffer tank (30); a second fluid communication system including valves (502) and communication means (501) between the valves (502), and each bioprocessing microfluidic device (20) is in fluid communication with each buffer tank (30); system.

2. Further comprising at least one waste liquid tank (42), the second fluid communication system includes valves (502) and communication means (501) between the valves (502), and each bioprocessing microfluidic device (20) is in fluid communication with each buffer tank (30) or the waste tank (42); the communication system includes two independent flow lines, a first flow line and a second flow line, the first flow line communicating a reservoir (40) with the microfluidic device (20) via a buffer tank (30), and the second flow line communicating each microfluidic device (20) and each buffer tank (30) with the waste tank (42); The system for processing biological particles according to claim 1 .

3. 2. The system for processing biological particles as described in claim 1, wherein each reservoir (40), each buffer tank (30), and each bioprocessing microfluidic device (20) further includes a waste tank (42) in fluid communication with the waste tank (42) via the first fluid communication system and / or the second fluid communication system.

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

5. 2. The system for processing biological particles of claim 1, wherein the valve is a chip (503) configured to open fluid communication when two chips (503) are in contact and to close fluid communication when the chip (503) is not in contact with another chip (503).

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

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

8. 2. The system for processing biological particles as described in claim 1, wherein the number of valves (502) of 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. 2. The system for processing biological particles of claim 1, wherein the number of valves (502) in the first and second fluid communication systems is less than the number of ports in all bioprocessing microfluidic devices (20) multiplied by the number of reservoirs (40).

10. 2. 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 of claim 1 , wherein the system includes at least two buffer tanks.

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

13. A method for processing biological particles using a system according to any one of claims 1 to 12, comprising: i. flowing a liquid containing bioparticles from at least one reservoir (40) through said first fluid communication system to at least one buffer tank (30); ii. Flowing a liquid containing biological particles from at least one buffer tank (30) through said second fluid communication system to at least one bioprocessing microfluidic device (20); A method comprising:

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