Material conveying device for in-vitro biological reaction and corresponding detection and reaction system

Abstract

The utility model provides a material conveying device for in-vitro biological reaction, which is used for circularly conveying materials in the in-vitro biological reaction and comprises a material input pipeline, a material output pipeline and a conveying unit, the conveying unit is respectively communicated with one end of the material input pipeline and one end of the material output pipeline, and the material output pipeline is communicated with the conveying unit. The conveying unit is used for driving the conveying of the materials; the other end of the input pipeline is communicated with a reactor for in-vitro biological reaction, so that the material from the reactor is conveyed to the conveying unit through the material input pipeline, and the other end of the material output pipeline is communicated with the reactor. The utility model also provides a corresponding detection and reaction system. The utility model provides a material conveying device for an in-vitro biological reaction, which is used for solving the problem that the activity of materials is reduced when the materials need to be circularly conveyed in the in-vitro biological reaction process.

Description

Technical Field

[0001] The utility model relates to the technical field of material conveying for in vitro biological reactions, in particular to a material conveying device for in vitro biological reactions and a corresponding detection and reaction system. Background Art

[0002] In vitro biological reactions, such as cell-free in vitro biosynthesis - obtaining the basic components required for transcription and translation in cells, adding DNA templates in vitro to maintain gene transcription, protein translation or metabolic processes, and synthesizing target products. Cell-free synthetic biology (CFSE) removes the cell membrane and can directly regulate the life activities inside the cell; removes the natural genome, eliminates non-essential gene regulation, and decouples cell growth and core metabolic regulation; the system is open, without material transport barriers, easy to add substrates, remove products and monitor and analyze the process, with the greatest degree of engineering freedom, playing an important role in basic disciplines and engineering applications: revealing life systems such as protein translation mechanisms; and has broad application potential in structural biology, high-throughput screening, biocatalysis, biomedicine and other fields.

[0003] In large-scale in vitro biological reaction processes, materials in the reaction process sometimes need to be extracted and circulated. For example, activity detection is performed during circulated transportation to monitor the quality of the reaction at any time. The activity of materials is easily affected by shear force. If the transportation method is improper, the activity of the materials circulated during the reaction will be reduced. When the cycle is continuous, there will be a greater impact, which will directly affect the stability of the material activity and the accuracy of the test results, further affect the control of the reaction process, and ultimately affect the yield of the reaction, and may also complicate the production process.

[0004] Currently, there is no research on the reduction in activity caused by improper delivery methods when cyclic delivery is required during in vitro biological reactions. Utility Model Content

[0005] The utility model provides a material conveying device for in vitro biological reaction, which can reduce the problem of activity reduction caused by cyclic conveying in in vitro biological reaction.

[0006] In order to solve the above technical problems, the utility model provides a material conveying device for in vitro biological reaction, which is used for circulating the material in the in vitro biological reaction, and includes: a material input pipeline, a material output pipeline, and a conveying unit, wherein the conveying unit is respectively connected with one end of the material input pipeline and one end of the material output pipeline, and the conveying unit is used to drive the conveying of the material; the other end of the input pipeline is connected with the reactor of the in vitro biological reaction so that the material from the reactor is conveyed to the conveying unit through the material input pipeline, and the other end of the material output pipeline is connected with the reactor.

[0007] Specifically, an activity detection unit is provided on the material output pipeline or the material input pipeline, and the material is transported to the activity detection unit for activity detection. Preferably, the activity detection unit includes: a detection pipeline and at least one detection sensor provided on the detection pipeline. Further preferably, the detection sensor is selected from one or more of a dissolved oxygen sensor, a pH sensor, a turbidity sensor, a conductivity sensor, a liquid concentration sensor, a liquid temperature sensor, a liquid flow sensor or an ethanol concentration sensor.

[0008] Specifically, the conveying unit includes: a buffer tank, a vacuum pump and an air pipe. The buffer tank is respectively connected to the material input pipeline, the material output pipeline, the vacuum pump and one end of the air pipe. The vacuum pump is used to evacuate the buffer tank to maintain a predetermined vacuum degree; the air pipe is used to input gas to allow the material in the buffer tank to be output from the material output pipeline. The vacuuming, the input gas, the material input and the material output are all controlled by separate valves.

[0009] Specifically, the buffer tank is connected to the material output pipeline and the material input pipeline through one material opening or two material openings, and the buffer tank is connected to the vacuum pump and the air pipe through one air opening or two air openings.

[0010] Specifically, a liquid level switch is provided on the inner wall of the buffer tank to control the maximum liquid level of the material in the buffer tank.

[0011] Specifically, the number of the buffer tanks is multiple, and the multiple buffer tanks are arranged in parallel, wherein the material input pipeline, the material output pipeline, the vacuum pump, and the air pipe are connected to the multiple buffer tanks through their own multiple pipelines, and the pipelines are each provided with a separate valve or

[0012] The material input pipeline, the material output pipeline, the vacuum pump, and the air pipe are connected to the plurality of buffer tanks through a main pipe and branch pipes, and separate valves are provided on the branch pipes.

[0013] Specifically, the buffer tank is further provided with a vacuum sensor. Preferably, the vacuum sensor is a field emission vacuum sensor or a piezoresistive vacuum sensor.

[0014] Specifically, the delivery unit is a reciprocating pump. Preferably, the reciprocating pump is selected from a plunger pump or a piston pump.

[0015] Specifically, when the activity detection unit is arranged in the material input pipeline, the material passes through the activity detection unit and then through the reciprocating pump; when the activity detection unit is arranged in the material output pipeline, the material passes through the reciprocating pump and then through the activity detection unit.

[0016] A second aspect of the utility model provides a detection system, comprising: a material conveying device and an activity detection unit as described in the above technical solution.

[0017] Specifically, the detection system further includes a control unit, and the control unit is communicatively connected with the activity detection unit and the material conveying device.

[0018] The third aspect of the utility model provides an in vitro biological reaction system, comprising: the detection system as described in the above technical solution and a reactor connected to the detection system.

[0019] Beneficial effects of the utility model:

[0020] (1) Solve the problem that when materials need to be circulated during in vitro biological reactions, for example, when materials are circulated for activity testing to monitor the quality of the reaction at any time, the activity of the materials is easily affected by shear force;

[0021] (2) Provide a material conveying device without mechanical contact with the material, meet the material circulation conveying requirements of in vitro biological reactions, and will not cause the activity of the material to decrease;

[0022] (3) A material conveying device with a low shear force pump is also provided to meet the material circulation conveying requirements of in vitro biological reactions, and the activity of the material is reduced within an acceptable range. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] In order to more clearly illustrate the technical solution of the utility model, the following briefly introduces the drawings required for use in the utility model. Obviously, the drawings described below are only some embodiments of the utility model. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0024] Figure 1 It is an embodiment of the utility model;

[0025] Figure 2 It is another embodiment of the utility model;

[0026] Figure 3 It is another embodiment of the utility model;

[0027] Figure 4 It is another embodiment of the utility model;

[0028] Figure 5 It is the influence on the fluorescence value when the delivery unit is a plunger pump;

[0029] Figure 6 It is the influence on the fluorescence value when the delivery unit is a gear pump;

[0030] Figure 7 It is the influence of the gear pump discharge on the fluorescence value;

[0031] Figure 8 It is the influence on the fluorescence value when the delivery unit is a diaphragm pump.

[0032] in, Figure 1-4 The figures in the figure are as follows: 1. material input pipeline, 2. material output pipeline, 3. buffer tank, 4. vacuum pump, 5. air pipe, 6. detection pipeline, 7. valve, 8. liquid level switch, 9. reciprocating pump. DETAILED DESCRIPTION

[0033] The following will be combined with the accompanying drawings to clearly and completely describe the technical solutions in the utility model. Obviously, the described embodiments are part of the embodiments of the utility model, not all of the embodiments. Based on the embodiments of the utility model, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the utility model.

[0034] In vitro biological reaction refers to the reaction of synthesizing the target organism in an in vitro cell-free system, such as in vitro protein synthesis, in vitro RNA synthesis, etc. In vitro protein synthesis includes but is not limited to IVT reaction (in vitro translation reaction), IVTT reaction (in vitro transcription and translation reaction), and IVDTT reaction (in vitro replication transcription and translation reaction). IVTT reaction is preferred in the embodiments of the present application. IVTT reaction, corresponding to the IVTT system, is the process of transcribing and translating DNA into protein in vitro.

[0035] Example 1

[0036] The utility model provides a material conveying device for an in vitro biological reaction, which is used for cyclically conveying materials in the in vitro biological reaction, and comprises: a material input pipeline 1, a material output pipeline 2, and a conveying unit, wherein the conveying unit is respectively connected with one end of the material input pipeline 1 and one end of the material output pipeline 2, and the conveying unit is used to drive the conveying of the material; the other end of the material input pipeline 1 is connected with the reactor of the in vitro biological reaction so that the material from the reactor is conveyed to the conveying unit through the material input pipeline 1, and the other end of the material output pipeline 2 is connected with the reactor.

[0037] like Figure 1-4 As shown, an activity detection unit is provided on the material output pipeline 2 or the material input pipeline 1, and the material is transported to the activity detection unit for activity detection. Specifically, the activity detection unit includes: a detection pipeline and at least one detection sensor provided on the detection pipeline 6. More specifically, the detection sensor is selected from one or more of a dissolved oxygen sensor, a pH value sensor, a turbidity sensor, a conductivity sensor, a liquid concentration sensor, a liquid temperature sensor, a liquid flow sensor or an ethanol concentration sensor. Specifically, the detection pipeline 6 is provided on the material output pipeline 2 or the material input pipeline 1, and both ends of the detection pipeline 6 are respectively connected to the material output pipeline 2 or the material input pipeline 1, and at least one socket is provided on the detection pipeline 6 for at least one detection sensor to be inserted to monitor the material condition during the reaction process. The detection pipeline 6 is provided with multiple sockets for multiple detection sensors to be inserted one by one to monitor the material condition during the reaction process.

[0038] like Figure 1 As shown, the conveying unit includes: a buffer tank 3, a vacuum pump 4 and an air pipe 5. The buffer tank 3 is respectively connected with one end of the material input pipeline 1, the material output pipeline 2, the vacuum pump 4 and the air pipe 5. The vacuum pump 4 is used to evacuate the buffer tank 3 to maintain a predetermined vacuum degree; the air pipe 5 is used to input gas to allow the material in the buffer tank 3 to be output from the material output pipeline 2. The vacuuming, the input gas, the material input and the material output are all controlled by a separate valve. Specifically, the buffer tank 3 is first evacuated to a predetermined vacuum degree by the vacuum pump 4, and then the material input pipeline 1 and the buffer tank 3 are connected by valve control. The material is pressed into the buffer tank 3 due to the vacuum degree in the buffer tank 3. After the material reaches the predetermined liquid level, the connection between the material input pipeline 1 and the buffer tank 3 is cut off by valve control, and then the air pipe 5 and the buffer tank 3 are connected by valve control. The air is input to the buffer tank 3 by valve control to output the material in the buffer tank 3 from the material output pipeline 2. This embodiment adopts a driving design scheme of vacuum suction and air pressure return, which is particularly suitable for the delivery of materials that are susceptible to fluid shear force in in vitro biological reactions. It can reduce or avoid the impact of material flow on the yield of in vitro biological reactions.

[0039] like Figure 1-2As shown, the buffer tank 3 is connected to the material output pipeline 2 and the material input pipeline 1 through one material opening or two material openings, and the buffer tank 3 is connected to the vacuum pump 4 and the air pipe 5 through one air opening or two air openings. Specifically, the buffer tank 3 can be any one of one material opening and one air opening, one material opening and two air openings, two material openings and one air opening, or two material openings and two air openings.

[0040] like Figure 1-2 As shown, a liquid level switch 8 is provided on the inner wall of the buffer tank 3 to control the maximum liquid level of the material in the buffer tank 3. Specifically, when the material is pressed into the buffer tank 3 due to the vacuum in the buffer tank 3, the predetermined liquid level reached by the material cannot exceed the maximum liquid level. Once the material reaches the maximum liquid level, the liquid level switch 8 is immediately activated to discharge the material from the buffer tank 3.

[0041] There are multiple buffer tanks 3, and multiple buffer tanks 3 are arranged in parallel, wherein, in one case: the material input pipeline 1, the material output pipeline 2, the vacuum pump 4, and the air pipe 5 are connected to the multiple buffer tanks 3 one-to-one through their respective multiple pipelines, and separate valves are provided on the pipelines, or in another case: the material input pipeline 1, the material output pipeline 2, the vacuum pump 4, and the air pipe 5 are connected to the multiple buffer tanks 3 through the main pipe and the branch pipes, and separate valves are provided on the branch pipes. Specifically, there are multiple pipelines on the material input pipeline 1 that are connected to the multiple buffer tanks 3 one-to-one, and the multiple pipelines can be in the form of a main pipe and a branch pipe, or can be multiple independent pipelines. The material output pipeline 2, the vacuum pump 4, the air pipe 5 and the material input pipeline 1 have the same aforementioned pipeline structure. For example, Figure 2 As shown, when two buffer tanks 3 are arranged in parallel, the two buffer tanks 3 are first evacuated to a predetermined vacuum degree through the branch pipes of the vacuum pump 4 and the connecting pipes of the two buffer tanks 3, and then the branch pipes are controlled by valves to connect the material input pipeline 1 and the first buffer tank 3. The material is pressed into the first buffer tank 3 due to the vacuum degree in the buffer tank 3. After the material reaches a predetermined liquid level, the connection between the material input pipeline 1 and the first buffer tank 3 is cut off through valve control. At this time, the air pipe 5 and the first buffer tank 3 are connected through valve control to connect the branch pipes of the two buffer tanks. The air is input to the first buffer tank 3 through valve control to output the material in the first buffer tank 3 from the material output pipeline 2. At the same time, the material input pipeline 1 and the second buffer tank 3 are connected through valve control to connect the branch pipes of the two buffer tanks. The material is pressed into the second buffer tank 3 due to the vacuum degree in the second buffer tank 3, that is, the first buffer tank 3 is discharging material while the second buffer tank 3 is taking in material. When the first buffer tank 2 has finished discharging material, it is adjusted alternately so that the first buffer tank 3 takes in material and the second buffer tank 3 takes out material. The same operation is performed when the number of buffer tanks 3 is greater than 2, and different buffer tanks 3 take turns to discharge and take in materials, so as to keep the material output pipeline 2 uninterrupted.

[0042] The buffer tank 3 is also provided with a vacuum sensor, and specifically, the vacuum sensor is a field emission vacuum sensor or a piezoresistive vacuum sensor. Specifically, the vacuum sensor can monitor the vacuum degree in the buffer tank 3 in real time and transmit a monitoring signal.

[0043] A second aspect of the utility model provides a detection system, comprising: a material conveying device and an activity detection unit as described in the above technical solution.

[0044] Specifically, the detection system also includes a control unit, which is communicatively connected with the activity detection unit and the material conveying device.

[0045] The third aspect of the utility model provides an in vitro biological reaction system, comprising: a detection system as described in the above technical solution and a reactor connected to the detection system.

[0046] Example 2

[0047] The part that this embodiment differs from the first embodiment is the conveying unit.

[0048] like Figure 3-4 As shown, the conveying unit is a reciprocating pump 9, and specifically, the reciprocating pump 9 is selected from a plunger pump or a piston pump. Specifically, the material conveying device for the in vitro biological reaction uses a reciprocating pump 9 such as a plunger pump or a piston pump, which can reduce or avoid the influence of the shear force of the pump on the biological activity during the material conveying process, and thus does not affect the output of the in vitro biological reaction. In the flow direction of the material, the reciprocating pump 9 can be set in front of or behind the activity detection unit.

[0049] In order to verify the problems found in this application and effectively solve them, the following experiments were conducted. In the following experimental examples, two groups of protein reaction systems with the same composition were used for reaction:

[0050] (1) Reaction system 1: During the reaction process, a pump is added to continuously circulate the reaction materials:

[0051] Experimental Example 1, Experimental Example 2 and Experimental Example 3: After 3 hours of reaction, the material was subjected to fluorescence detection (pump group, fluorescence detection one); after 3 hours of reaction, the material was discharged by pump, and the discharged material was subjected to fluorescence detection (pump discharge group, fluorescence detection two);

[0052] (2) Reaction system 2: No transport is performed during the reaction:

[0053] Experimental Example 1, Experimental Example 2 and Experimental Example 3: After 3 hours of reaction, the material was subjected to fluorescence detection (PC group, fluorescence detection three); after 3 hours of reaction, the material in Example 1 and Example 3 was discharged by a pump, and the discharged material was subjected to fluorescence detection (PC discharge group, fluorescence detection four).

[0054] Experimental Example 1

[0055] This experimental example 1 uses embodiment 2 for testing.

[0056] The reciprocating pump 9 used in the conveying unit is a plunger pump. The plunger pump group is the reaction system 1 that uses the plunger pump to circulate and convey the material. The PC group is the reaction system 2. The above-mentioned fluorescence detection is performed on the two reaction systems after 3 hours of reaction. The results are as follows: Figure 5 As shown:

[0057] (1) The plunger pump group (fluorescence detection 1) reduced the fluorescence value of the IVTT reaction solution by approximately 5% compared to the PC group (fluorescence detection 3);

[0058] (2) There is no significant change in the fluorescence value of the IVTT reaction liquid in the PC discharge group (fluorescence detection four) compared with the PC group (fluorescence detection three), and there is no significant change in the fluorescence value of the IVTT reaction liquid in the plunger pump discharge group (fluorescence detection two) compared with the plunger pump group (fluorescence detection one).

[0059] This experimental example 1 shows that the material is limitedly affected by the circulating transport of the plunger pump, reducing the synthetic yield of the product by about 5% (the reaction is a protein production reaction, the protein is fluorescent, and the fluorescence value represents the protein yield). Within an acceptable range, the plunger pump can be used for circulating material delivery.

[0060] Experimental Example 2

[0061] The difference between Experimental Example 2 and Experimental Example 1 is the conveying unit. The conveying unit of Experimental Example 2 uses a gear pump. The gear pump group uses the gear pump to circulate the material in the reaction system 1 and the PC group reaction system 2. Fluorescence detection is performed on the two reaction systems after 3 hours of reaction. The results are as follows: Figure 6-7 As shown:

[0062] (1) Figure 6 As shown, the gear pump group (fluorescence detection one) reduced the fluorescence value of the IVTT reaction solution by approximately 25% compared to the PC group (fluorescence detection three);

[0063] (2) Figure 7 As shown, the gear pump discharge group (fluorescence detection 2) does not have obvious changes in the fluorescence value of the IVTT reaction liquid compared to the gear pump group (fluorescence detection 1).

[0064] This experimental example 2 shows that the material is easily affected by the gear pump's cyclic delivery, thereby reducing the synthetic yield of the product by about 25% (the reaction is a protein production reaction, the protein is fluorescent, and the fluorescence value represents the protein yield); however, for non-cyclic delivery, it will not be affected by the gear pump delivery. Therefore, the researchers found that in the in vitro biological reaction process, such as in vitro protein synthesis and in vitro RNA synthesis, the cyclic delivery will cause activity loss. When the delivery needs to maintain activity during the reaction process (for example, for activity detection), the delivery method in this process needs to be changed to reduce the shear force.

[0065] Experimental Example 3

[0066] The difference between Experimental Example 2 and Experimental Example 1 is the delivery unit. The delivery unit of Experimental Example 2 uses a diaphragm pump. The diaphragm pump group is the reaction system 1 that uses the diaphragm pump to circulate and deliver materials, and the PC group is the reaction system 2. The above-mentioned fluorescence detection is performed on the two reaction systems after 3 hours of reaction. The results are as follows Figure 8 As shown:

[0067] (1) The diaphragm pump group (fluorescence detection 1) reduced the fluorescence value of the IVTT reaction solution by approximately 35% compared to the PC group (fluorescence detection 3);

[0068] (2) There is no significant change in the fluorescence value of the IVTT reaction liquid in the PC discharge group (fluorescence detection four) compared with the PC group (fluorescence detection three), and there is no significant change in the fluorescence value of the IVTT reaction liquid in the diaphragm pump discharge group (fluorescence detection two) compared with the diaphragm pump group (fluorescence detection one).

[0069] The comparative example of Experiment 3 shows that the material will be affected by the diaphragm pump circulation, thereby reducing the synthetic yield of the product by about 35%; while for non-circulation transportation, it will not be affected by the gear pump transportation. It is generally believed that the diaphragm pump is a low shear pump, and the shear force effect brought by the diaphragm pump should be small. However, the effect of the diaphragm pump circulation in this experimental example is greater than that of the gear pump circulation in Experiment 2. The reason is speculated that the diaphragm pump generally has a diaphragm and a screen, and the shear force effect brought by the two to the IVTT reaction material may be greater.

[0070] It can be seen from the above experimental examples that for in vitro biological reactions that require cyclic transportation, the cyclic transportation method should try to avoid the influence of shear force, so a pump that reduces shear force or a power method that has no mechanical contact with the material can be used to transport the material; and the output of the product after the reaction is completed will not be significantly affected by the shear force.

[0071] The above description is only a preferred embodiment of the present invention and is not intended to limit the scope of protection of the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A material conveying device for in vitro biological reaction, used for circulating and conveying materials in in vitro biological reaction, characterized in that: include: Material input pipeline, material output pipeline, conveying unit, among which, The conveying unit is respectively connected to one end of the material input pipeline and one end of the material output pipeline, and the conveying unit is used to drive the conveying of the material; The other end of the input pipeline is connected to the reactor of the in vitro biological reaction so that the material from the reactor is transported to the transport unit through the material input pipeline, and the other end of the material output pipeline is connected to the reactor.

2. The material conveying device according to claim 1, characterized in that: in, The material output pipeline or the material input pipeline is provided with an activity detection unit, and the material is transported to the activity detection unit for activity detection.

3. The material conveying device according to claim 1 or 2, characterized in that: in, The conveying unit includes: a buffer tank, a vacuum pump and an air pipe. The buffer tank is respectively connected to one end of the material input pipeline, the material output pipeline, the vacuum pump and the air pipe. The vacuum pump is used to evacuate the buffer tank to maintain a predetermined vacuum degree; The air pipe is used to input gas so that the material in the buffer tank can be output from the material output pipeline. The vacuuming, the gas input, the material input and the material output are all controlled by separate valves.

4. The material conveying device according to claim 3, characterized in that: in, The buffer tank is connected to the material output pipeline and the material input pipeline through one material opening or two material openings. The buffer tank is connected with the vacuum pump and the air pipe through one air opening or two air openings.

5. The material conveying device according to claim 3, characterized in that: A liquid level switch is provided on the inner wall of the buffer tank to control the maximum liquid level of the material in the buffer tank.

6. The material conveying device according to claim 3, characterized in that: The number of the buffer tanks is multiple, and the multiple buffer tanks are arranged in parallel, wherein: The material input pipeline, the material output pipeline, the vacuum pump, and the air pipe are connected to the plurality of buffer tanks through respective pipelines, and the pipelines are provided with separate valves or The material input pipeline, the material output pipeline, the vacuum pump, and the air pipe are connected to the plurality of buffer tanks through a main pipe and branch pipes, and separate valves are provided on the branch pipes.

7. The material conveying device according to claim 3, characterized in that: The buffer tank is also provided with a vacuum sensor.

8. The material conveying device according to claim 1 or 2, characterized in that: The conveying unit is a reciprocating pump.

9. The material conveying device according to claim 2, characterized in that: The activity detection unit includes: a detection pipeline and at least one detection sensor arranged on the detection pipeline.

10. The material conveying device according to claim 9, characterized in that: The detection sensor is selected from one or more of a dissolved oxygen sensor, a pH sensor, a turbidity sensor, a conductivity sensor, a liquid concentration sensor, a liquid temperature sensor, a liquid flow sensor or an ethanol concentration sensor.

11. The material conveying device according to claim 7, characterized in that: The vacuum sensor is a field emission vacuum sensor or a piezoresistive vacuum sensor.

12. The material conveying device according to claim 8, characterized in that: The reciprocating pump is selected from a plunger pump or a piston pump.

13. A detection system, characterized in that: include: A material conveying device and an activity detection unit as described in any one of claims 1 to 12.

14. The detection system according to claim 13, characterized in that: It also includes a control unit, which is communicatively connected with the activity detection unit and the material conveying device.

15. An in vitro bioreactor system, characterized in that: include: A detection system as claimed in claim 13 or 14 and a reactor connected to the detection system.

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