BIOPROCESSING DEVICE

Abstract

A system for processing biological particles including bioprocessing microfluidic devices, reservoirs, buffer tanks and two fluidic connection systems. A first fluidic connection system includes valves and connecting elements between valves, so that each reservoir or port configured to connect a reservoir may be in fluidic connection with each buffer tank, and a second fluidic connection system includes valves and connecting elements between valves, so that each bioprocessing microfluidic device may be in fluidic connection with each buffer tank.

Claims

1.-12. (canceled)

13. A system for processing biological particles comprising: i. at least four bioprocessing microfluidic devices; ii. at least three reservoirs or ports configured to connect a reservoir; iii. at least one buffer tank; and iv. at least two fluidic connection systems; wherein a first fluidic connection system comprises valves and connecting means between valves, so that each reservoir or port configured to connect a reservoir may be in fluidic connection with each buffer tank; and wherein a second fluidic connection system comprises valves and connecting means between valves, so that each bioprocessing microfluidic device may be in fluidic connection with each buffer tank.

14. The system for processing biological particles according to claim 13, further comprising a waste tank, so that each reservoir, each buffer tank and each bioprocessing microfluidic device may be in fluidic connection with waste tank through the first fluidic connection system and/or through the second fluidic connection system.

15. The system for processing biological particles according to claim 13, wherein connecting means comprise tubes.

16. The system for processing biological particles according to claim 13, wherein valves are tips configured to open fluidic connection when two tips are in contact and configured to close fluidic connection when a tip is not in contact with another tip.

17. The system for processing biological particles according to claim 13, wherein the inner volume of the second connection system is less than 300% of volume of all bioprocessing microfluidic devices.

18. The system for processing biological particles according to claim 13, wherein the number of valves of the first fluidic connection system is less than the number of reservoirs multiplied by three times the number of buffer tanks.

19. The system for processing biological particles according to claim 13, wherein the number of valves of the second fluidic connection system is less than the number of ports of all bioprocessing microfluidic devices multiplied by the number of buffer tanks.

20. The system for processing biological particles according to claim 13, wherein the number of valves of the first and second fluidic connection systems is less than the number of ports of all bioprocessing microfluidic devices multiplied by the number of reservoirs.

21. The system for processing biological particles according to claim 13, wherein buffer tanks are controlled by a pressure source.

22. The system for processing biological particles according to claim 13, wherein system comprises at least two buffer tanks.

23. The system for processing biological particles according to claim 13, wherein the microfluidic devices are enclosed in a pressurized chamber.

24. Method for processing biological particles using a system according to claim 13, the method comprising: i. flowing liquid containing biological particles from at least one reservoir into at least one buffer tank through the first fluidic connection system; and ii. flowing liquid containing biological particles from at least one buffer tank into at least one bioprocessing microfluidic device through the second fluidic connection system. iii.

25. The system for processing biological particles according to claim 13, wherein biological particles are biological cells.

26. The method for processing biological particles according to claim 24, wherein biological particles are biological cells.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0053] Features and advantages of the invention will become apparent from the following description of embodiments of a system and a method according to the disclosure, this description being given merely by way of example and with reference to the appended drawings in which:

[0054] FIG. 1 is a schematic architecture of a system for processing biological particles in a fixed topology configuration of connecting means.

[0055] FIG. 2 is a schematic architecture of a system for processing biological particles with a part of connecting means in a fixed topology and another part of connecting means in a dynamic topology.

[0056] FIG. 3 is a schematic architecture of a system for processing biological particles in which dynamic topology of connecting means is achieved by moving microfluidic devices.

ILLUSTRATIVE EMBODIMENTS OF THE INVENTION

[0057] FIG. 1 shows a system (1) according to a first embodiment of the disclosure, intended to process biological particles. Six microfluidic devices (20) are placed in a chamber (2) of the system (1). Each microfluidic device comprises an inlet and an outlet (i.e. two ports), both ending with a valve (502). Ten reservoirs (40) are placed in the system (1) and comprise an outlet ending with a valve (502). Here, reservoirs (40) are refrigerated in a refrigerated chamber (4). Four buffer tanks (30) are placed in the system (1) and comprise an inlet/outlet ending with a valve (502). Here, buffer tanks (30) are temperature controlled in a chamber (3), typically at the temperature biological cells are processed. Between valves (502) are arranged connecting means (501) in the form of tubes. By proper configuration of open and closed valves, each reservoir can be in fluidic connection with each buffer tank and each buffer tank can be in fluidic connection with each microfluidic device.

[0058] In the present application, a buffer tank is a fluidic element in which liquid is introduced, temporarily stored, then drained out. Buffer tanks may be chambers or elongated tubes for instance.

[0059] Here, the first fluidic connection system comprises valves (502) associated to reservoirs (40) and buffers tanks (30) and connecting means (501) between these valves (502). 28 valves (502) are used to connect 10 reservoirs (40) with 4 buffer tanks (30). The second fluidic connection system comprises valves (502) associated to microfluidic devices (20) and buffers tanks (30) and connecting means (501) between these valves (502). Valves (502) associated with buffer tanks (30) are part of both the first and the second fluidic connection systems.

[0060] Microfluidic devices (20) are further linked to control modules (22, 23) for temperature and dissolved gas concentration in chamber (2). Water content of microfluidic devices is further controlled by a module (24) to measure water loss and eventually add or remove water in microfluidic devices if required. When water loss is caused by evaporation, water vapor is added in the chamber comprising the microfluidic devices (20).

[0061] As illustrated in a non-limitative way, system (1) comprises a waste tank (42), which may be in fluidic connection with each reservoir (40), each buffer tank (30) and each microfluidic device (20). In the illustrated embodiment, the set of connecting means (501) located closest to reservoirs (40), buffer tanks (30) and microfluidic devices (20) (via inlet) is used to flow content of reservoirs (40) into microfluidic devices (20) through temporary storage in buffer tanks (30), defining a first flow line. The set of connecting means (501) located farthest of reservoirs (40), buffer tanks (30) and microfluidic devices (20) (via outlet) is used to flow liquids into the waste (42), defining a second flow line. With such configuration, liquid disposal to the waste (42) does not use the same connecting means (501) as liquid delivery to the microfluidic channels (20).

[0062] Besides, in the illustrated embodiment the connection systems comprise two independent flow lines connecting microfluidic devices (20) to buffer tanks (30) or to the waste tank (42). With such a configuration, it is possible to flow a liquid from a first buffer tank (30) so as to harvest the content of a microfluidic device (20), this content being transferred simultaneously in a second buffer tank (30) as the volume of the microfluidic device remains essentially constant. Having at least two buffer tanks (30) which may be connected via different flow lines to a single microfluidic device (20) allows recollecting liquids from a microfluidic device, in particular when it comprises a product or biological particles of interest, for their transfer to an outer container connected via a port or into a reservoir (40).

[0063] In the example shown in FIG. 1, buffer tanks (30) are controlled by a pressure source (311), here a pressure controller. By depression of the pressure source (311), liquid flow is induced from reservoir (40) or microfluidic device (20) into a buffer tank (30). With increased pressure of the pressure source (311), liquid flow is induced from buffer tank (30) to microfluidic device (20), reservoir (40) or waste (42). To avoid formation of bubbles induced by low pressure, it is preferred to use an increased pressure of the pressure source. In the specific case of a flow imposed from a first buffer tank (30) to a microfluidic device (20) to harvest the content of said microfluidic device (20) into a second buffer tank (30), the first buffer tank (30) is pressurized and the second buffer tank (30) is kept at a pressure high enough to avoid bubble formation.

[0064] According to this embodiment of the system (1), a controller (10) with a user interface (11) and a central computer (101) enables setting flows in the system according to the bioprocess considered. The controller monitors parameters: temperature, pressure, humidity, gas concentration in microfluidic devices, water loss of microfluidic devices, time and duration of process steps and defines flows between all components of the system in terms of flow rates and displaced volumes.

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

[0066] This variant is typically obtained by addition of a fluidic connection between two sub-systems, each sub-system being illustrated in FIG. 1. For instance, one reservoir (40) may be substituted by the fluidic connection between both sub-systems.

[0067] With this variant, versatility of the system is increased. Bioprocessing microfluidic devices may be stored at different temperatures, while using the same reservoirs. Several reservoir chambers may be also controlled at different temperatures, depending on the chemicals stored. Several buffers may be used for specific steps of liquid flow, avoiding cross contamination. Last but not least, this parallelization variant allows to increase on demand the number of bioprocessing microfluidic devices used in similar conditions.

[0068] FIG. 2 shows a system (1) according to a second embodiment of the disclosure, intended to process biological particles. Elements similar to those of the first embodiment bear identical references. Six microfluidic devices (20) are placed in a chamber (2) of the system (1). Each microfluidic device comprises two ports: an inlet tip (505) and an outlet tip (505) acting as valves. Ten reservoirs (40) are placed in the system (1) and comprise a tip (505) acting as a valve. Here, reservoirs (40) are refrigerated in a refrigerated chamber (4). Four buffer tanks (30) are placed in the system (1) and comprise an inlet/outlet ending with a valve (502). Here, buffer tanks (30) are temperature controlled in a chamber (3), typically at the temperature biological cells are processed. Between valves (502) are arranged connecting means (501) in the form of tubes. Two tips (505) acting as valves are arranged on tubes inside two injectors (506). The first fluidic connection system comprises valves (502), injectors (506) and tips (505) associated with buffer tanks (30) and tips (505) associated with reservoirs and connecting means (501) between these valves/tips. The second fluidic connection system comprises valves (502), injectors (506) and tips (505) associated with buffer tanks (30) and tips (505) associated with microfluidic devices (20) and connecting means (501) between these valves/tips.

[0069] As illustrated in a non-limitative way, buffer tanks (30) and pressure source (311) are placed on a moving head (510), whose displacement is controlled by an arm (511). By proper move of the moving head (510), the tip (505) of one injector (506) is brought in contact with a tip (505) of a reservoir (40), thus opening a fluidic connection of the first fluidic connection system. Then, after another move of the moving head (510), the tip (505) of one injector (506) is brought in contact with a tip (505) of a microfluidic device (20), thus opening a fluidic connection of the second fluidic connection system.

[0070] In the example shown in FIG. 2, two fluidic connections are realized simultaneously between a microfluidic device (20) and the two injectors (506). One fluidic connection is used to flow liquid from a first buffer tank (30) to microfluidic device (20) and the second fluidic connection is used to flow liquid from microfluidic device (20) to a second buffer tank (30), thus keeping the volume of microfluidic device (20) constant. Liquid removed from microfluidic device (20) is stored in buffer tank (30) and may be discarded to waste (42) or used further in bioprocesses in another microfluidic device (20) or stored in a reservoir (40) as a final product.

[0071] In this embodiment, the volume of connecting means (501) is very strongly limited, as the topology of connecting means (501) is dynamically adapted on demand with displacement of moving head (510). In particular the inner volume of the second connection system is not dependent of the number of microfluidic devices (20) nor of the distance between them. Thus, volumes transferred from a buffer tank (30) to a microfluidic device is almost totally transferred without liquid remaining in dead portions. In addition, only 20 valves/tips are used to connect 10 reservoirs with 4 buffer tanks. And 22 valves/tips are used to connect 6 microfluidic devices with two ports each with 4 buffer tanks. A total of 32 valves/tips is sufficient to connect 10 reservoirs and 6 microfluidic devices with a great versatility of flows and processes.

[0072] In this embodiment, the chamber (2) is pressurized so that pressure in the chamber (2) is higher than pressure in microfluidic devices (20). This overpressure avoids any risk of leak through the tips (505). When two tips (505) are in contact, the high pressure or low pressure generated by the pressure source (311) is sufficient to flow liquid through the tips (505).

[0073] FIG. 3 shows a system (1) according to a third embodiment of the disclosure, intended to process biological particles. Elements similar to those of the first and second embodiments bear identical references. twelve microfluidic devices (20) are placed in a chamber (2) of the system (1). Each microfluidic device comprises two ports: an inlet tip (505) and an outlet tip (505) acting as valves. Ten reservoirs (40) are placed in the system (1) and comprise an outlet ending with a valve (502). Here, reservoirs (40) are refrigerated in a refrigerated chamber (4). Four buffer tanks (30) are placed in the system (1) and comprise an inlet/outlet ending with a valve (502). Here, buffer tanks (30) are temperature controlled in a chamber (3), typically at the temperature biological cells are processed. Between valves (502) are arranged connecting means (501) in the form of tubes. Two tips (505) acting as valves are arranged on tubes inside two injectors (506) which are fixed. The first fluidic connection system comprises valves (502) associated with buffer tanks (30) and reservoirs, injectors (506) and tips (505) and connecting means (501) between these valves/tips. The second fluidic connection system comprises valves (502), injectors (506) and tips (505) associated with buffer tanks (30) and tips (505) associated with microfluidic devices (20) and connecting means (501) between these valves/tips.

[0074] As illustrated in a non-limitative way, a moving head (510), whose displacement is controlled by an arm (511), can hold and move a microfluidic device (20) in different locations in the chamber (1). By proper move of the moving head (510), the tips (505) of both injectors (506) are brought in contact with two tips (505) of the microfluidic device (20), thus opening a fluidic connection of the second fluidic connection system, similarly as in the second embodiment. Injectors (506) may be mounted on mechanical actuators and/or be equipped with detectors such as contact or pressure sensors allowing to adjust the coupling with feedback. The topology of the second fluidic connection system is adapted dynamically on demand. On the other hand, the first fluidic connection system is similar to the first embodiment. This embodiment is particularly relevant when high numbers of microfluidic devices (20) are used, for example more than 100, while few reservoirs (40) are used in the system (1).

[0075] In the example shown in FIG. 3, an additional bioprocessing module (7) is arranged in the chamber. This additional module can be used for selective processes on one microfluidic device (20) moved by the moving head (510), such as washing, cell sorting (optically, magnetically or by size exclusion), electroporation, filtration, lysis, microinjection, purification, ion exchange or any usual bioprocessing step such as amplification, concentration, purification, gene editing, gene delivery, RNA delivery, protein delivery, differentiation, dedifferentiation, harvest, sorting of cells, and harvest and purification.

[0076] In the example shown in FIG. 3, an additional analysis module (8) is arranged in the system (1). A microfluidic device (20) may be moved in the analysis module (8) by the moving head (510), then the microfluidic device (20) itself or the liquid contained therein is 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 usual analysis protocol. Specifically, a microfluidic device (20) may be used as low volume reservoir for transfer to the analysis module (8) and then analysis. This microfluidic device (20) may be configured for specific analysis.