Static reactor and its use in biochemical reactions

Abstract

The application provides a static reactor, which comprises a first storage chamber, a second storage chamber and a reaction chamber; each structure is separated by a film structure and is provided with a sample inlet, and a sealing cover is arranged on the sample inlet; a trigger switch is arranged to start the reaction; an output switch is arranged to obtain the reaction product; and the bottom surface of the reaction chamber is provided in an uneven structure. The static reactor has reasonable structural design, high automation degree, convenient operation and use, and does not need dynamic mixing. The application of the static reactor in biochemical reactions and the application of the static reactor in a cell-free protein synthesis method are also provided.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, and more specifically to a static reactor, which can be used for solid-liquid phase biochemical reactions, and a method for in vitro cell-free protein synthesis using the static reactor. Background Technology

[0002] In biochemical reactions, to ensure a more complete and efficient reaction, it is usually necessary to continuously and thoroughly mix the components in the reaction system. Effective mixing enables effective collisions between reactant molecules, which in turn allow the biochemical reaction to proceed. The frequency of these effective collisions directly affects the reaction rate. Brownian motion of molecules can contribute to effective collisions within the reaction system. To increase the frequency of these collisions, external mixing forces are typically used in larger reaction systems to intensify molecular motion and enhance effective collisions. Common external mixing methods include mechanical stirring, magnetic stirring, and shaking. In single-phase reaction systems, such as those in solution, the components are in the same solvent and can move freely, allowing for sufficient and effective collisions to occur. In multiphase reaction systems, such as those involving insoluble or slightly soluble solids reacting with solutions, the limited contact area between the solid and solution, coupled with the tendency for solid sedimentation and aggregation to significantly reduce the effective contact area, necessitates external mixing forces to prevent sedimentation and aggregation, ensuring the reaction proceeds fully and effectively. External mixing typically requires additional mixing devices, such as mechanical stirrers, magnetic stirrers, shakers, and rollers, which increases costs and occupies experimental space. Therefore, static reactors that do not require dynamic mixing but achieve reaction efficiency comparable to continuous external mixing represent the future trend in biochemical reactors. These static reactors are easy to use, require no additional mixers, and effectively reduce instrument costs and operating space. Summary of the Invention

[0003] This invention provides a static reactor with a reasonable structural design, high degree of automation, convenient operation, and no need for dynamic mixing, to solve the problem of the need for continuous mixing and shaking of the reaction system during the reaction process. This reactor is particularly suitable for solid-liquid phase reactions.

[0004] To achieve the above objectives, the present invention provides the following technical solution: a static reactor comprising two storage chambers, one reaction chamber, and two control switches (i.e., a trigger switch and an output switch). Both storage chambers are located above the reaction chamber, and the upper and lower storage chambers, the lower storage chamber and the reaction chamber, and the reaction chamber and the outlet are separated by a thin film with a thickness of micrometers. The bottom surfaces of both the upper storage chamber and the reaction chamber are designed as inclined surfaces with a certain angle to reduce liquid phase retention. The bottom surface of the reaction chamber is configured with an uneven structure, preferably a triangular prism array, to enhance the retention and dispersion of solids during the reaction process and reduce the aggregation of solids. The additional retention and dispersion surface provided by the triangular prism array effectively increases the contact area between the solid and liquid phases during the reaction process, thereby increasing the reaction efficiency. Both the upper and lower storage chambers and the reaction chamber have their own inlets and sealing caps. The central portion of the sealing caps of the upper storage chamber and the reaction chamber is a thin film with a thickness of micrometers. The membrane directly beneath the sealing cap of the upper storage chamber corresponds to the membrane between the upper and lower storage chambers, and between the lower storage chamber and the reaction chamber. This means the control switch (trigger switch) on the storage chamber side can sequentially puncture the membranes of the upper storage chamber sealing cap, the upper and lower storage chambers, and the lower storage chamber and the reaction chamber. Similarly, the membrane directly beneath the sealing cap of the reaction chamber corresponds to the membrane between the reaction chamber and the outlet. This means the control switch (output switch) on the reaction chamber side can sequentially puncture the membrane of the reaction chamber sealing cap and the membrane between the reaction chamber and the outlet. Above the sealing cap of the upper storage chamber is the reaction trigger switch, a pointed design used to simultaneously puncture the three membranes: the upper storage chamber sealing cap, the membrane between the upper and lower storage chambers, and the membrane between the lower storage chamber and the reaction chamber. This allows the upper storage chamber to connect to the atmosphere, while simultaneously allowing liquid samples from both storage chambers to flow into the reaction chamber, thus triggering the reaction. Above the sealing cap of the reaction chamber is the reaction output switch, a pointed design used to puncture the reaction chamber sealing cap and the two membranes between the reaction chamber and the outlet, allowing the reactants in the reaction chamber to flow out through the outlet. An additional thin vent tube connects the upper storage chamber to the reaction chamber to balance the air pressure, allowing the liquid sample in the storage chamber to flow smoothly into the reaction chamber after the reaction trigger switch is turned on.

[0005] Specifically, a first aspect of the present invention provides a static reactor, the reactor comprising:

[0006] The system comprises a first storage chamber, a second storage chamber, and a reaction chamber. The second storage chamber is located below the first storage chamber and is separated by a first thin-film structure. The reaction chamber is located below the second storage chamber and is separated by a second thin-film structure. A third thin-film structure is provided at the outlet of the reaction chamber. Each of the first, second, and reaction chambers has a sample inlet with a sealing cap. The sealing caps of the sample inlets of the first and reaction chambers are respectively provided with thin-film structures. A trigger switch is provided at the sample inlet of the first storage chamber to puncture the thin-film structure, the first thin-film structure, and the second thin-film structure on the corresponding sealing cap. An output switch is provided at the sample inlet of the reaction chamber to puncture the thin-film structure and the third thin-film structure on the corresponding sealing cap. The bottom surface of the reaction chamber is designed to be uneven.

[0007] Furthermore, the first storage chamber, the second storage chamber, and the reaction chamber can be integrally formed into a reactor by the outer shell or assembled into a reactor by separate molding. If integral molding technology is adopted, it can be integrally molded by 3D printing technology, which is simple to manufacture and effectively ensures that the reagents stored inside the reactor are not contaminated by the outside when the reaction is not turned on.

[0008] Furthermore, the trigger switch and output switch body can be of any shape, such as a cylinder or prism, preferably a hexagonal prism, and the switch end has a pointed structure to facilitate puncturing the film and to facilitate mixing of reactants through the gap between its body and the damaged film.

[0009] Furthermore, the bottom surfaces of both the first storage chamber and the reaction chamber are designed with a small-angle inclination, ranging from 2 to 15° relative to the horizontal plane, preferably 5 to 10°, so as to minimize liquid residue when the liquid sample flows out of the storage chamber and the reaction material flows out of the reaction chamber.

[0010] Furthermore, the injection port and sealing cap can be connected by any of the following methods or combinations: snap-fit ​​structure, threaded structure, or vacuum grease seal.

[0011] Furthermore, a baffle with aperture is installed on the outlet passage to block punctured membranes, etc. The baffle can also be replaced with a filter screen or filter cloth with filtering function according to the requirements of the reaction, so as to filter the reaction products.

[0012] Furthermore, the uneven structure is a triangular prism array, and at least one inclined surface of the bottom surface of the reaction chamber is designed as an uneven structure. Preferably, the bottom surface of the reaction chamber has three inclined surfaces, and a triangular prism array is designed and constructed on each inclined surface. The passage from the reaction chamber to the third thin film structure is designed as a constricted structure to ensure that the mixture in the reaction chamber can be completely discharged and prevent accumulation at the corners. In the above-mentioned triangular prism array, the base triangle of each triangular prism is an isosceles right triangle with a hypotenuse of 1 mm, and the interval between each triangular prism is 0.2 mm.

[0013] Furthermore, the reactor also includes legs, which are positioned on one side of the uneven structure. The legs are of fixed length or adjustable length, thereby concentrating the reactants in the area where the triangular prism array is located. For example, the legs can be set in two sections, and the length of the legs can be manually adjusted, or the legs can be a nested structure, and the length of the legs can be manually adjusted through nesting.

[0014] Furthermore, the reactor also includes an internal pressure balancing device. For example, an additional venting tube can be connected between the first storage chamber and the reaction chamber to balance the pressure between the reaction chamber and the upper storage chamber, allowing the liquid sample to flow smoothly into the reaction chamber.

[0015] Furthermore, the thickness of the film is 1 to 500 μm, preferably 5 μm, 10 μm, 15 μm, 20 μm, 30 μm, 40 μm, 50 μm, 60 μm, 70 μm, 80 μm, 90 μm, 100 μm, 200 μm, 300 μm, 400 μm, or 500 μm.

[0016] Furthermore, the total volume of the reactor can be any size, and the volumes of the first storage chamber, the second storage chamber, and the reaction chamber can be any size, such as the volume of the first storage chamber being 0.5 mL to 50 mL, the volume of the second storage chamber being 50 μL to 1000 μL, and the volume of the reaction chamber being 2 mL to 80 mL.

[0017] Furthermore, there are no special restrictions on the reactor material; it can be prepared from a variety of different types of materials, such as polydimethylsiloxane (PDMS), 3D printing materials, glass, paper-based materials, and plastics.

[0018] A second aspect of the invention provides the application of any one or any combination of the static reactors described in the first aspect in biochemical reactions.

[0019] A third aspect of the present invention provides a method for in vitro cell-free protein synthesis, comprising the steps of:

[0020] (1) Provide a static reactor of any one or any combination thereof in the first aspect;

[0021] Place the same or different buffer solutions in the first storage chamber;

[0022] Place a liquid containing DNA and / or RNA, which may be the same or different, into the second storage chamber; and

[0023] Place the same or different biological reaction raw materials in the reaction chamber;

[0024] (2) Activate the trigger switch to allow the buffer solution and liquid containing DNA and / or RNA to flow into the corresponding reaction chamber and mix with the biological reaction raw materials in the reaction chamber to form a reaction system and carry out the reaction;

[0025] (3) After the reaction is complete, turn on the output switch to allow the liquid containing the same or different proteins to flow out, and obtain the liquid containing the synthesized target protein.

[0026] Furthermore, the bioreactor feedstock is a lyophilized powder containing cell extracts.

[0027] The main advantages and positive effects of this invention include:

[0028] (1) This invention is a static reactor, which does not require additional mixing equipment, such as mechanical stirrers, magnetic stirrers, shaking tables, rollers, etc., which effectively reduces the cost of instruments and operating space;

[0029] (2) The reactor does not require additional internal circuits, external controllers or other professional equipment. It is simple to operate, convenient to use, low in cost, and does not require professional operators, making it easy to promote and popularize.

[0030] (3) The reactor can be integrally formed by 3D printing and other technologies, which is simple to manufacture and effectively ensures that the reagents stored inside the reactor are not contaminated by the outside when the reaction is not turned on; or it can be assembled after being formed separately.

[0031] (4) The reactor has a wide range of applications, not only applicable to biochemical reactions, but also has no special restrictions on reactor materials. It can be prepared from a variety of different types of materials, such as polydimethylsiloxane (PDMS), 3D printing materials, glass, paper-based materials, plastics, etc., and the specific materials can be determined artificially according to the application field of the reactor and the characteristics of the reactants.

[0032] (5) The reactor of this application also has an internal pressure balancing device (such as a venting tube), which can allow the liquid to flow smoothly while reducing the amount of outside air introduced, thereby reducing the contamination of the reagent. Attached Figure Description

[0033] Figure 1 Left view of the static reactor

[0034] Figure 2 Static reactor front sectional view

[0035] Figure 3 Top view of the static reactor

[0036] Figure 4 Left view of the static reactor

[0037] Figure 5Results of relative fluorescence unit determination experiment

[0038] In the attached figure, the reference numerals are as follows:

[0039] 1 is the first storage chamber, 2 is the second storage chamber, 3 is the reaction chamber, 4 is the first thin film structure, 5 is the second thin film structure, 6 is the third thin film structure, 7 is the outlet, 8 is the inclined surface of the bottom of the first storage chamber, 9 is the inclined surface of the bottom of the reaction chamber, 10 is the triangular prism array, 11 is the sample inlet of the first storage chamber, 12 is the sample inlet of the second storage chamber, 13 is the sample inlet of the reaction chamber, 14 is the first sealing cap, 15 is the second sealing cap, 16 is the wedge end of the first sealing cap, 17 is the wedge end of the second sealing cap, 18 is the first circular thin film structure, 19 is the second circular thin film structure, 20 is the third sealing cap, 21 is the wedge end of the third sealing cap, 22 is the trigger switch, 23 is the pointed structure of the trigger switch, 24 is the main body of the trigger switch, 25 is the output switch, 26 is the pointed structure of the output switch, 27 is the main body of the output switch, 28 is the venting capillary tube, and 29 is the leg. Detailed Implementation

[0040] It should be understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be described in detail here.

[0041] In this invention, the cell extract does not contain intact cells. Typical yeast cell extracts include ribosomes for protein translation, transfer RNA, aminoacyl-tRNA synthetase, initiation and elongation factors required for protein synthesis, and termination release factors. Furthermore, yeast extracts also contain other proteins derived from the cytoplasm of yeast cells, particularly soluble proteins.

[0042] like Figure 1 , 2 As shown in Figures 3 and 4, the present invention provides a static reactor, which includes an outer shell, and a first storage chamber 1, a second storage chamber 2 and a reaction chamber 3 are fixed inside the outer shell. The overall structure including the outer shell can be integrally formed by molding technology such as 3D printing. Of course, alternatively, the first storage chamber 1, the second storage chamber 2 and the reaction chamber 3 can be separately formed and then assembled to form the reactor.

[0043] The first storage chamber 1 (also called the upper storage chamber) is a liquid sample storage chamber, and the second storage chamber 2 (also called the lower storage chamber) is a liquid sample storage chamber. The upper and lower storage chambers are separated by a first thin film structure 4 (circular film) with a thickness of 300 micrometers. This film is fixed inside the device and completely blocks the upper and lower storage chambers. The reaction chamber 3 is used to store solid samples and is separated from the lower storage chamber 2 and the outlet 7 by a second thin film structure 5 and a third thin film structure 6 (which adopt the same design as the first thin film structure 4), respectively. The bottom surfaces of the first storage chamber 1 and the reaction chamber 3 are both designed with a small-angle inclined surface, with an angle range of 2 to 15° relative to the horizontal plane, preferably 5 to 10°, so as to minimize liquid residue when the liquid sample flows out of the upper storage chamber and the reaction material flows out of the reaction chamber. The inclination angles of the inclined surface 8 of the bottom surface of the first storage chamber and the inclined surface 9 of the bottom surface of the reaction chamber can be the same or different. At least one inclined surface 9 of the bottom surface of the reaction chamber is designed as an uneven structure, preferably a triangular prism array 10. Preferably, the bottom surface of the reaction chamber is provided with three inclined surfaces. Figure 3 Each inclined surface is designed with a triangular prism array 10. The passage from the reaction chamber to the third thin film structure 6 is designed with a constricted structure to ensure that the mixture in the reaction chamber can be completely discharged and prevent accumulation at the corners. The base triangle of each triangular prism in the above triangular prism array 10 is an isosceles right triangle with a hypotenuse of 1 mm, and the interval between each triangular prism is 0.2 mm. The first storage chamber 1, the second storage chamber 2 and the reaction chamber 3 all have sample inlets. The sample inlet 12 of the second storage chamber 2 is located on the back of the reactor; the sample inlet 11 of the first storage chamber 1 and the sample inlet 13 of the reaction chamber 3 are of the same specification, and the sample inlets are all located at the top of the chamber.

[0044] The inlet 11 of the first storage chamber 1 and the inlet 13 of the reaction chamber 3 are each equipped with a first sealing cap 14 and a second sealing cap 15. The wedge end 16 of the first sealing cap 14 and the wedge end 17 of the second sealing cap 15 are designed as frustum shapes with a certain angle to fit tightly into each inlet. With a small amount of vacuum grease, a good sealing effect can be achieved. The center of the first sealing cap 14 and the second sealing cap 15 contains a first circular thin film structure 18 and a second circular thin film structure 19 with a thickness of 300 micrometers. After the two sealing caps are sealed, the first circular thin film structure 18 is located directly above the first thin film structure 4 and the second thin film structure 5, and the second circular thin film structure 19 is located directly above the third thin film structure 6.

[0045] The sample inlet 12 of the second storage chamber 2 is equipped with a third sealing cap 20. The wedge end 21 of the third sealing cap 20 is designed as a frustum with a certain angle to fit tightly into the sample inlet 12. It is equipped with a small amount of vacuum grease to achieve a good sealing effect. When not in use, the first storage chamber, the second storage chamber, and the reaction chamber are in a sealed and mutually isolated state.

[0046] Above the injection port 11 is a reaction trigger switch 22 (i.e., the first puncture structure). The end of the trigger switch has a pointed structure 23. The main body 24 of the trigger switch 22 can be of any shape, such as a cylinder or prism, preferably a hexagonal prism. By pressing the trigger switch 22 with a finger, the first circular thin film structure 18, the first thin film structure 4, and the second thin film structure 5 are punctured, allowing the reaction liquid in the upper and lower storage chambers to flow into the reaction chamber 3 simultaneously through the gaps between the main body 24 of the trigger switch 22 and each thin film structure. This mixture mixes with the solid sample in the reaction chamber 3, thus initiating the reaction. The specifications of the main body 24 of the trigger switch 22 can be determined manually, thereby controlling the distance between the main body 24 and the flow channel, and thus controlling the overall reaction time or the reaction rate of each step.

[0047] Above the inlet 13 is the reaction output switch 25 (i.e., the second puncture structure). The end of the output switch 25 has a pointed structure 26, and the main body 27 of the output switch can be any shape, such as a cylinder or prism, preferably a hexagonal prism. Pressing with a finger triggers the output switch 25, simultaneously puncturing the second circular membrane structure 19 and the third membrane structure 6, allowing the mixture in the reaction chamber 3 to exit from the outlet 7, thus terminating and exiting the reaction. A baffle with pores is installed in the passage of the outlet 7 to block the punctured membranes, etc. The baffle can also be replaced with a filter screen or filter cloth with a filtering function, depending on the requirements of the reaction, thereby filtering the reaction products.

[0048] In addition, optionally, an internal pressure balancing device can be provided to allow the sample to flow more smoothly into the reaction chamber. For example, an additional venting tube 28 can be connected between the first storage chamber 1 and the reaction chamber 3 to balance the pressure between the reaction chamber and the upper storage chamber, so that the liquid sample can flow smoothly into the reaction chamber.

[0049] Alternatively, the three injection ports and their corresponding sealing caps can be connected by a snap-fit ​​structure, allowing the sealing caps to be opened and closed by rotating and moving them. The use of a small amount of vacuum grease can also achieve a complete sealing effect for the device. Alternatively, the injection ports and their corresponding sealing caps can be connected by a threaded structure.

[0050] When using a static reactor, the reactor can be tilted toward the side with the triangular prism array 10 to facilitate better contact between the reactants and the triangular prism array 10. The reactants can also be concentrated in the area where the triangular prism array 10 is located by setting the legs 29 of the reactor to be telescopic or adjustable in length. For example, the legs 29 can be set in two sections so that the length of the legs can be manually adjusted, or the legs 29 can be a nested structure so that the length of the legs can be manually adjusted by nesting.

[0051] Example 1

[0052] Taking a cell-free in vitro protein synthesis system as an example, the specific usage method of the static reactor of this application is introduced. This is a verification example and does not mean that the reactor can only be used in a cell-free in vitro protein synthesis system. The reaction reagents used are as follows:

[0053] Biochemical lyophilized powder: The mixture before lyophilization contains 50% by volume yeast cell extract and 50% by volume buffer.

[0054] Buffer solution:

[0055] 10-50 mM phosphate buffer, 10-100 mM tris(hydroxymethyl)aminomethane, 20-300 mM potassium acetate, 10-100 mM magnesium acetate, 1-20 mM dithiothreitol, 0.1%-5% polyethylene glycol 8000, 2-50 mM glucose, 10-200 mg / mL dextrin, 0.001-0.01 mg / mL amylase, 0.1-30 mM nucleoside triphosphate mixture, and 0.08-0.24 mM amino acid mixture.

[0056] DNA: Green fluorescent protein DNA.

[0057] Add 4 mL of deionized water to the first storage chamber 1 through the inlet 11 of the static reactor, and then cover it with the first sealing cap 14.

[0058] 1 g of lyophilized biochemical powder is added to reaction chamber 3 through inlet 13 on the reactor, and the second sealing cap 15 is placed on top, ensuring that the lyophilized biochemical powder is in full contact with the triangular prism array 10 on the inclined surface 9 of the bottom of the reaction chamber.

[0059] Add 170 µL of liquid containing green fluorescent protein DNA (DNA concentration 10 ng / µL) to the second storage chamber 2 through the inlet 12 on the reactor, and then cover it with the third sealing cap 20.

[0060] Pressing the reaction trigger switch 22 simultaneously destroys the first circular thin film structure 18 on the first sealing cover 14 of the first storage chamber 1 in the reactor, the first thin film structure 4 between the first storage chamber 1 and the second storage chamber 2, and the second thin film structure 5 between the second storage chamber 2 and the reaction chamber 3, allowing deionized water and DNA to flow into the reaction chamber 3, mix with the biochemical freeze-dried powder in the reaction chamber 3, shake well, let stand, and react at room temperature.

[0061] After 2-6 hours, press the reaction output switch 25 to simultaneously break the membrane structure on the sealing cover of the reaction chamber in the reaction device unit, as well as the membrane structure between the reaction chamber and the outlet, so that the protein-containing liquid flows out, and a liquid containing green fluorescent protein is obtained. Immediately take 10 µL and place it in a 384-well plate, centrifuge at 4000 rpm for 1 minute, and use an enzyme-linked immunosorbent assay (ELISA) reader to measure the RFU value (RFU, Relative Fluorescence Unit).

[0062] Comparative Example 1

[0063] Using the same batch of reaction reagents as in Example 1, mix them into a 15 mL centrifuge tube and place the centrifuge tube on a mixer. Perform continuous mixing reaction at the same time and place as in Example 1. After the reaction time is the same as in Example 1, immediately take 10 µL and place it in a 384-well plate. Centrifuge at 4000 rpm for 1 minute and measure the RFU value using a microplate reader.

[0064] Comparative Example 2

[0065] A reactor without the triangular prism array 10 is used, and the rest of the reactor is completely identical in structure to the reactor in Example 1.

[0066] Using the same reaction reagents as in Example 1, and loading them into the reactor of Comparative Example 2 in the same manner, the reaction trigger switch 22 was pressed, the mixture was shaken and allowed to stand. The reaction was carried out under the same reaction conditions (room temperature) as in Example 1.

[0067] After the reaction time is the same as in Example 1, press the reaction output switch 25 to simultaneously break the membrane structure on the sealing cover of the reaction chamber in the reaction device unit and the membrane structure between the reaction chamber and the outlet, so that the protein-containing liquid flows out and a liquid containing green fluorescent protein is obtained. Immediately take 10 µL and place it in a 384-well plate, centrifuge at 4000 rpm for 1 minute, and use an enzyme-linked immunosorbent assay (ELISA) reader to determine the RFU value.

[0068] like Figure 5 As shown, the relative fluorescence units represent the amount of green fluorescent protein synthesized in the reaction system. D and E are two replicates of Example 1, C is the experimental result of Comparative Example 1, and A and B are two replicates of Comparative Example 2. It can be seen from the results that the relative fluorescence units in Comparative Example 2 are only 50% to 65% of those in Example 1, while the relative fluorescence units in Example 1 are close to or higher than those in Comparative Example 1. This shows that the reactor with the triangular prism array can achieve or exceed the effect of a centrifuge tube in a continuously mixed state, and the relative fluorescence units are more than twice as high as those in the reactor without the triangular prism array.

[0069] Combination Figure 5The experimental results demonstrate that the triangular prism array 10 on the bottom surface of reaction chamber 3 can effectively improve the static reaction rate of this type of biochemical reaction, achieving a reaction efficiency comparable to that of continuous external mixing without the need for dynamic mixing. This type of static reactor does not require additional mixing equipment such as mechanical stirrers, magnetic stirrers, shakers, or rollers, effectively reducing the user's instrument costs and operating space requirements.

[0070] All documents mentioned in this invention are incorporated herein by reference as if each document were individually incorporated by reference. Furthermore, it should be understood that after reading the foregoing teachings of this invention, those skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by the appended claims.

Claims

1. A static reactor, the reactor comprising: First storage chamber (1), second storage chamber (2), and reaction chamber (3); The second storage chamber (2) is located below the first storage chamber (1) and is separated by the first membrane structure (4); the reaction chamber (3) is located below the second storage chamber (2) and is separated by the second membrane structure (5); a third membrane structure (6) is provided at the outlet (7) of the reaction chamber (3); The first storage chamber (1), the second storage chamber (2) and the reaction chamber (3) are all equipped with sample inlets, and the sample inlets are equipped with sealing caps. The sealing caps of the sample inlets of the first storage chamber (1) and the reaction chamber (3) are respectively equipped with thin film structures. The sample inlet (11) of the first storage chamber (1) is equipped with a trigger switch (22) to puncture the thin film structure, the first thin film structure (4) and the second thin film structure (5) on the sealing cap corresponding to the sample inlet (11); An output switch (25) is provided at the injection port (13) of the reaction chamber (3) to puncture the thin film structure and the third thin film structure (6) on the sealing cap corresponding to the injection port (13); The bottom surfaces of the first storage chamber (1) and the reaction chamber (3) are both designed with a small-angle inclination, with the angle ranging from 2 to 15° relative to the horizontal plane. The bottom surface of the reaction chamber is set as an uneven structure; the uneven structure is a triangular prism array (10); the bottom surface of the reaction chamber is provided with three inclined surfaces, and a triangular prism array is designed and constructed on each inclined surface.

2. The static reactor according to claim 1, characterized in that: The first storage chamber (1), the second storage chamber (2), and the reaction chamber (3) are formed as a reactor by integral molding of the outer shell or by assembly of separate molding parts.

3. The static reactor according to claim 1, characterized in that: The main body of the trigger switch (22) and the output switch (25) can be of any shape, and their ends are pointed.

4. The static reactor according to claim 1, characterized in that: The injection port and sealing cap are connected by any of the following methods or combinations: snap-fit ​​structure, threaded structure, or vacuum grease seal.

5. The static reactor according to claim 1, characterized in that: A baffle, filter screen, or filter cloth with pore size is provided on the passage where the outlet (7) is located.

6. The static reactor according to any one of claims 1-5, characterized in that: It also includes legs (29), which are set on one side of the uneven structure and are of fixed length or adjustable length.

7. The static reactor according to any one of claims 1-5, characterized in that: It also includes an internal air pressure balancing device.

8. The application of any static reactor of claims 1-7 in biochemical reactions.

9. A method for in vitro cell-free protein synthesis, characterized in that, Including the following steps: a: Provide a static reactor as described in any one of claims 1-7; Place the same or different buffer solutions in the first storage chamber (1); Place a liquid containing DNA and / or RNA, which may be the same or different, into the second storage chamber (2); and Place the same or different biological reaction raw materials into the reaction chamber (3); b: Activate the trigger switch (22) to allow the buffer solution and liquid containing DNA and / or RNA to flow into the corresponding reaction chamber (3) and mix with the biological reaction raw materials in the reaction chamber (3) to form a reaction system and carry out the reaction; c: After the reaction is complete, start the output switch (25) to allow the liquid containing the same or different proteins to flow out, and obtain the liquid containing the synthesized target protein.

10. The method according to claim 9, wherein the bioreaction raw material is a lyophilized powder containing cell extracts.

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