Bioreactor and method for biological detection based on the same

Abstract

The present invention relates to the field of biotechnology, and in particular to a bioreactor and a method for performing biological detection based on the device. The present invention discloses a bioreactor for biomacromolecules, comprising a power module 1, a control module 2, a liquid processing module 3, a reactor module 4 and a sensor 5; the power module 1 comprises a DC power supply 6 and a switch 7; the control module 2 comprises a system controller 8, an input device 9 and an output device 10; the liquid processing module 3 comprises a valve 11 or a combination of the valves 11, a pump 12 or a combination of the pumps 12 and a sample pool 13; the reactor module 4 comprises a reactor 14; the reactor 14 comprises a reactor frame 15 and a reactor cavity 16 formed by the reactor frame 15. The device has a simple and practical structure and is easy to operate. In the reactor, a uniform and highly sensitive reaction can be achieved using a very small amount of reagents.

Description

Technical Field

[0001] The present invention relates to the field of biotechnology, and in particular to a bioreactor and a method for performing biodetection based on the bioreactor. Background Art

[0002] Common detection methods for biomacromolecules are all based on the structure of biomacromolecules and detection reagents to form feedback signals.

[0003] Common experimental methods are as follows

[0004] 1. Protein detection

[0005] A) Place the gel or other carrier containing the protein into the staining solution

[0006] B) Vibration and heating are used to promote the binding of dyes to proteins

[0007] C) Place the stained gel into the destaining solution

[0008] D) Vibration and heating are used to remove dyes that are not bound to proteins from the gel.

[0009] 2. Nucleic acid testing

[0010] A) Place the nucleic acid gel or other carrier into the staining solution

[0011] B) Vibration and heating are used to promote the binding of dyes to nucleic acids

[0012] C) Place the stained gel into the destaining solution

[0013] D) Vibration, heating, etc. are used to remove dyes that are not bound to nucleic acids from the gel.

[0014] 3. Probe detection

[0015] A) Place the carrier with the sample in the blocking solution

[0016] B) Accelerate the binding of unbound sample areas to the sealant by vibration and heating

[0017] C) Place the blocked carrier into the probe solution

[0018] D) Accelerate the combination of probe and sample through vibration and heating

[0019] E) Place the carrier in the wash

[0020] F) Wash away non-specifically bound probes by shaking and heating

[0021] The general detection method of biomacromolecules can be described as the immersion method. Through immersion in a large amount of solution, the molecules are subjected to specific binding and non-specific removal by relying on the thermal motion of the molecules. Although the thermal motion of the molecules can be accelerated by vibration, heating, ultrasound, microwaves, etc., it still cannot avoid its existing defects, such as time-consuming and labor-intensive, wasteful of reagents, poor repeatability, and long uniformity.

[0022] Therefore, providing an automated bioreactor for detecting biomacromolecules has important practical significance. Summary of the invention

[0023] In view of this, the present invention provides a biological reaction device and a biological detection method based on the device. The device has a simple and practical structure and is easy to operate. In the reactor, a uniform and highly sensitive reaction can be achieved using a very small amount of reagents.

[0024] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0025] The present invention provides a bioreactor, comprising a power module 1, a control module 2, a liquid processing module 3, a reactor module 4 and a sensor 5;

[0026] The power module 1 includes a DC power supply 6 and a switch 7;

[0027] The control module 2 includes a system controller 8, an input device 9 and an output device 10;

[0028] The liquid processing module 3 includes a valve 11 or a combination of the valves 11, a pump 12 or a combination of the pumps 12 and a sample pool 13;

[0029] The reactor module 4 includes a reactor 14; the reactor 14 includes a reactor frame 15 and a reactor cavity 16 formed by the reactor frame 15;

[0030] The power module 1 is connected to the control module 2, the liquid processing module 3, and the reactor module 4 respectively through cables 25;

[0031] The control module 2 is connected to the liquid processing module 3 and the reactor module 4 respectively through cables 25;

[0032] The liquid treatment module 3 is connected to the reactor module 4 via a pipeline 26;

[0033] The control module 2 is connected to the reactor module 4 via the sensor 5 .

[0034] In some specific embodiments of the present invention, the reactor frame 15 is provided with at least one opening 17 ; the opening 17 is connected to the pump 12 or a combination of the pumps 12 .

[0035] In some specific embodiments of the present invention, the reactor frame 15 is further provided with a pressing component 18 .

[0036] In some specific embodiments of the present invention, the reactor frame 15 is further provided with a sealing component 19 .

[0037] In some specific embodiments of the present invention, the reactor frame 15 includes a reactor front plate 20 and a reactor rear plate 21, and a limiting component 22 is further provided between the reactor front plate 20 and the reactor rear plate 21. The function of the limiting component is to prevent 96 and 15 from having an excessively large stroke during the pressing process, resulting in an excessively small volume of 16.

[0038] In some specific embodiments of the present invention, a mother liquid tank 23 is further provided between the liquid treatment module 3 and the reactor module 4 .

[0039] In some specific embodiments of the present invention, at least one flow dividing baffle 24 is further disposed in the reactor chamber 16 .

[0040] Based on the above research, the present invention also provides the application of the bioreactor in biological sample detection.

[0041] The present invention also provides a method for detecting biological samples. Based on the bioreactor provided by the present invention, a carrier carrying a substance to be reacted is placed in the reactor cavity 16, a reaction liquid is placed in the sample pool 13 of the liquid processing module 3, and a control module 2 is started by a power module 1 to control a pump 12 of the liquid processing module 3 or a combination of the pumps 12 to transport the reaction liquid in the sample pool (13) to the reactor cavity 16 to mix with the carrier carrying the substance to be reacted, react, and detect.

[0042] The present invention provides an automated bioreactor, which can automatically complete the entire process by only placing samples and reaction solutions at designated locations and setting the program.

[0043] Experiments show that:

[0044] In the Coomassie Brilliant Blue staining test, on IgG, all bands of the traditional method and the present invention are visible, but the present invention is more deeply colored. On BSA, the traditional method can see the 10th sample, and the present invention can see all 12 samples, and the present invention has better sensitivity. On lysozyme, the traditional method can see the 9th sample, and the present invention can see the 11th sample, and the present invention has better sensitivity. It can be seen from Figure 9 (C), Figure 9 (D), Figure 9 (E) and Tables 2 to 4 that compared with the traditional method, the coloring degree of the IgG test results of the present invention is increased by 4.49% to 22.90%; the coloring degree of the BSA test results is increased by 14.87% to 60.00%; the coloring degree of the lysozyme test results is increased by 9.72% to 28.37%. Compared with the traditional method, the present invention only needs to use 50% of the reagents, and the sensitivity and coloring depth are better than the traditional method.

[0045] In the silver staining test, when using half as much reagent, the sensitivity of the present invention is better than that of the traditional method, and the background is better than that of the traditional method. On IgG, all bands of the traditional method and the present invention are visible. On BSA, the 10th sample can be seen by the traditional method, and all 12 samples of the present invention are visible, and the sensitivity of the present invention is better. It can be seen from the results of Figure 10 (C), Figure 10 (D) and Tables 6 to 7 that compared with the traditional method, the coloring degree of the IgG test results provided by the method provided by the present invention is increased by 114.91% to 139.27%; the coloring degree of the BSA test results is increased by 100.88% to 186.47%. Compared with the traditional method, the present invention only needs to use 50% of the reagents, and its sensitivity and coloring degree are better than the traditional method.

[0046] In the WB1 test, at 93kd, the conventional method can be seen in the 4th lane, while the present invention can be seen in the 5th lane, indicating that the present invention has better sensitivity. At 7kd, the conventional method can be seen in the 8th lane, while the present invention can be seen in the 9th lane, indicating that the present invention has better sensitivity. As shown in Figure 11(E), Figure 11(F) and Tables 9 and 10, compared with the conventional method, the coloring degree of the 93kd detection result of the method provided by the present invention is increased by 102.09% to 507.93%; the coloring degree of the 7kd detection result is increased by 20.61% to 432.91%. Compared with the conventional method, the present invention only needs to use 74% of the reagents and 80% of the antibodies, and the sensitivity and signal strength of the present invention are better than those of the conventional method.

[0047] In the WB2 test, the conventional method can be seen in the 6th lane, while the present invention can be seen in the 8th lane, and the sensitivity of the present invention is better. As shown in Figure 12 (C) and Table 12, the color degree of the detection result of β-actin is improved by 3.05% to 305.33% compared with the conventional method. Compared with the conventional method, the present invention only needs to use 65% of the reagents, 40% of the antibodies and a shorter processing time. The sensitivity and signal strength of the present invention are better than those of the conventional method.

[0048] In the WB5 test, the colors of Figure 13 (B) and Figure 13 (D) are darker than those of Figure 13 (A) and Figure 13 (C). As shown in Figure 13 (E) and Table 13, the coloring degree of the GAPDH detection result of the method provided by the present invention is increased by 50.38% to 108.84% compared with the traditional method. That is, its signal intensity is greater, further indicating that the effect of the present invention is better than that of the traditional method. Compared with the traditional method, the present invention only needs to use 66% of the reagents, and the sensitivity and signal intensity of the present invention are better than those of the traditional method.

[0049] In the WB3 test, as can be seen from FIG14, the conventional method can see the 4th lane, while the present invention can see the 5th lane, and the sensitivity of the present invention is better. As can be seen from FIG14(C) and Table 15, the coloring degree of the β-actin detection result of the method provided by the present invention is improved by 27.52% to 603.57% compared with the conventional method. Compared with the conventional method, the present invention only needs to use 30% of the reagents and a shorter processing time, and the sensitivity and signal strength of the present invention are better than the conventional method.

[0050] In the WB4 test, it can be seen from Figure 15 (D) and Table 16 that the band depth of the present invention is stronger than that of the traditional method, and the coloring degree is increased by 59.26% to 111.13%, indicating that the present invention has a better effect. Compared with the traditional method, the present invention only needs to use 20% of the reagents and a shorter processing time, and the signal intensity of the present invention is better than that of the traditional method.

[0051] In summary, compared with traditional methods, this automated bioreactor has the following advantages:

[0052] 1. Save time. The system is fully automatic, saving a lot of manual work and greatly improving efficiency.

[0053] 2. High sensitivity. Through liquid flow control, the reaction efficiency is improved compared with traditional methods, and thus the sensitivity is higher under the same conditions as traditional methods.

[0054] 3. Save reactor reagents. In this automated bioreactor, the reactor only needs a small amount of liquid to complete the reaction of a large amount of reagents in the traditional method. BRIEF DESCRIPTION OF THE DRAWINGS

[0055] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art are briefly introduced below.

[0056] Figure 1 1 is a schematic diagram showing the structure of a bioreactor provided in Example 1 of the present invention;

[0057] Figure 22 is a schematic structural diagram of the bioreactor disclosed in Example 2 of the present invention;

[0058] Figure 3 2 is a schematic structural diagram of the bioreactor disclosed in Example 3 of the present invention;

[0059] Figure 4 14 shows the flow direction of the liquid in the reactor 14 on the time axis;

[0060] FIG. 5 is a schematic diagram showing the structure of the bioreactor disclosed in Examples 4 to 6 of the present invention; wherein FIG. 5(A) is a schematic diagram showing the structure of the bioreactor disclosed in Example 4 of the present invention; Figure 5(B) to Figure 5(D) FIG5(B) is a front view; FIG5(C) is a cross-sectional view; and FIG5(D) is a rear view. Figure 5(E) to Figure 5(G) FIG5(E) is a front view; FIG5(F) is a cross-sectional view; and FIG5(G) is a rear view.

[0061] FIG6 is a schematic diagram showing the structure of the bioreactor disclosed in Example 7 of the present invention; wherein FIG6(A) is a front view; FIG6(B) and FIG6(C) are side views showing the suspension of the reactant by the liquid;

[0062] Figure 7 Schematic diagram of the structure of the bioreactor disclosed in Example 8 of the present invention;

[0063] FIG8 is a schematic diagram of the structure of the bioreactor disclosed in Example 9 of the present invention; the bioreactor comprises a bidirectional quick plug; wherein FIG8(A) is a schematic diagram of the bidirectional quick plug; FIG8(B) is a schematic diagram of the quick plug B113; FIG8(C) is a cross-sectional view of the quick plug B113; FIG8(D) is a schematic diagram of the quick plug A112; and FIG8(E) is a cross-sectional view of the quick plug A112;

[0064] FIG9 shows the staining results of Example 10; wherein FIG9(A) shows the staining results of the conventional method; FIG9(B) shows the staining results of the present invention; FIG9(C) shows a comparative analysis of the IgG coloring degree in the staining results of the conventional method and the staining results of the present invention; FIG9(D) shows a comparative analysis of the lysozyme coloring degree in the staining results of the conventional method and the staining results of the present invention; FIG9(E) shows a comparative analysis of the BSA coloring degree in the staining results of the conventional method and the staining results of the present invention;

[0065] FIG10 shows the staining results of Example 11; FIG10(A) shows the staining results of the present invention; FIG10(B) shows the staining results of the conventional method; FIG10(C) shows a comparative analysis of the IgG staining degree in the staining results of the conventional method and the staining results of the present invention; FIG10(D) shows a comparative analysis of the BSA staining degree in the staining results of the conventional method and the staining results of the present invention;

[0066] FIG11 shows the results of Example 12; wherein FIG11(A) to FIG11(B) show the results of the conventional method; FIG11(C) to FIG11(D) show the results of the method provided by the present invention; FIG11(E) shows a comparative analysis of the coloring degree of the 93kd protein in the conventional method results and the results of the present invention; FIG11(F) shows a comparative analysis of the coloring degree of the 7kd protein in the conventional method results and the results of the present invention;

[0067] FIG12 shows the results of Example 13; FIG12(A) shows the results of the conventional method; FIG12(B) shows the results of the present invention; FIG12(C) shows a comparative analysis of the β-actin protein staining degree in the conventional method results and the results of the present invention;

[0068] FIG13 shows the results of Example 16; FIG13(A) shows the results of membrane A; FIG13(B) shows the results of membrane B; FIG13(C) shows the results of membrane C; FIG13(D) shows the results of membrane D; FIG13(E) shows a comparative analysis of the coloring degree of GAPDH protein in the results of the conventional method and the results of the present invention;

[0069] FIG14 shows the results of Example 14; FIG14(A) shows the results of the conventional method; FIG14(B) shows the results of the present invention; FIG14(C) shows a comparative analysis of the β-actin protein staining degree in the conventional method results and the present invention results;

[0070] FIG15 shows the results of Example 15; FIG15(A) shows the results of the conventional method; FIG15(B) shows the results of the present invention; FIG15(C) shows the flow direction of the primary antibody and secondary antibody mother solutions added to the corresponding reservoirs; FIG15(D) shows a comparative analysis of the coloring degree of the 7 kd protein in the conventional method results and the present invention results;

[0071] Among them, 1-power module; 2-control module; 3-liquid processing module; 4-reactor module; 5-sensor; 6-DC power supply; 7-switch; 8-system controller; 9-input device; 10-output device; 11-valve; 12-pump; 13-sample pool; 14-reactor; 15-reactor frame; 16-reactor cavity; 17-opening; 18-pressing component; 19-sealing component; 20-reactor front plate; 21-reactor rear plate; 22-limiting component; 23-mother liquor pool; 24-diversion block; 25-cable; 26-pipeline;

[0072] 17-1: liquid inlet; 17-2: liquid outlet; 27-waste liquid collector; 28-liquid level schematic component; 29-reaction liquid inlet and outlet pipeline; 30-waste liquid discharge or air inlet pipeline; 50, 51, 52, 53, 54, 55-two-position three-way valve; 32, 33, 34-a two-position three-way valve with three liquid ports, 33 is the common end, 32 and 33 are normally open; 38, 39, 40-a two-position three-way valve with three liquid ports, 39 is the common end, 38 and 39 are normally open Normally open connection; 35, 36, 37 are three liquid connection ports of a two-position three-way valve, 36 is the common end, 35 and 36 are normally open connection; 41, 42, 43 - three liquid connection ports of a two-position three-way valve, 43 is the common end, 42 and 43 are normally open connection; 44, 45, 46 - three liquid connection ports of a two-position three-way valve, 44 is the common end, 46 and 44 are normally open connection; 47, 48, 49 - three liquid connection ports of a two-position three-way valve, 49 is the common end, 47 and 49 are normally open connection;

[0073] 62-gas buffer bottle; 66, 67, 68, 69, 70, 71, 72, 73-eight-position valve selection interface; 74-common interface; 65-selection path, which connects the selection interface 66, 67, 68, 69, 70, 71, 72, 73 with the common interface 74 according to the signal of the control module 2; 64-four-position selection valve group;

[0074] 18-1: compression structure before the reactor, 18-2: compression structure after the reactor;

[0075] 91 - a rotating shaft of the pressing member, through which the pressing member 18 is fixed to the reactor frame 15 and can rotate around it;

[0076] 96 - reactor cover; 99 - connecting shaft between reactor cover 96 and reactor frame 15;

[0077] 106 - inner wall of reactor 14; 107 - reactants; 109 - schematic diagram of liquid flow;

[0078] 112-quick plug-in A; 113-quick plug-in B; 114-quick plug-in B113 pagoda connector, which is a derivative of 125 and is used to connect pipelines or directly connect to the container wall; 125-quick plug-in B113 closure, which forms a cavity with 116; 118-114 internal flow channel; 115-quick plug-in B113 internal pagoda pattern, used for internal leakage prevention and anti-falling after the quick plug-in A and the quick plug-in B are assembled; 116-quick plug-in B113 body; 11 7-conducting part; 119-internal flow channel of the conducting part; 120-guiding structure of the quick plug-in B113, which is used for internal guiding and fixing after being combined with the quick plug-in A; 121-limiting structure of the conducting part, which prevents the conducting part 117 from falling off from the quick plug-in B113; 122-internal limiting structure of the quick plug-in B113, which limits the inner retreat distance of the conducting part 117; 124-external limiting structure of the quick plug-in B113, which limits the outer retreat distance of the conducting part 117; 123-structure with one-way valve function The control plug-in B113 is separated from the plug-in A112, and the plug-in B is closed from the direction 118 to 119 and connected from the direction 119 to 118; 129-the upper closing structure of the plug-in A112, which is used to fix the anti-leakage ring 130; 130-the anti-leakage ring of the plug-in A112; 128-the main body of the plug-in A112; 127-the lower closing structure of the plug-in A112; 126-the pagoda joint of the plug-in A112; 131-the plug-in A112 2 guide structure; 132-126 internal flow channel; 133-a structural part with a one-way valve function, which controls the quick plug-in B113 to be closed from the direction of 132 to 129 and connected from the direction of 129 to 132 when the quick plug-in A112 is separated; when the quick plug-in A and the quick plug-in B are combined, the guide member 119 retreats to 122, opens 123 of 113, and makes it lose the one-way valve function, and 119 is inserted into 123 of 112, and makes it lose the one-way valve function. DETAILED DESCRIPTION

[0079] The present invention discloses a bioreactor and a method for performing biological detection based on the bioreactor. Those skilled in the art can refer to the content of this article and appropriately improve the process parameters to achieve the same. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art and are considered to be included in the present invention. The methods and applications of the present invention have been described through preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the methods and applications described herein without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0080] The purpose of the present invention is to provide an automated system for biomacromolecule reactions in view of the deficiencies in the prior art.

[0081] Another object of the present invention is to provide a method for detecting biomacromolecules using a biomacromolecule automated reaction system.

[0082] The purpose of the present invention can be achieved through the following technical solutions:

[0083] The present invention protects a biomacromolecule automated reaction system (in the present invention, it may also be referred to as a bioreactor), comprising a power module 1, a control module 2, a liquid processing module 3, a reactor module 4 and a sensor 5; the power module 1 provides driving energy and energy required for the reaction to the liquid processing module 3 and the reactor module 4, the liquid processing module 3 is controlled by the control module 2 and provides feedback of the operating status thereto, the liquid processing module 3 inputs or outputs reactants to the reactor module 4, the reactor module 4 provides feedback of the operating status to the control module 2 and the liquid processing module 3, and the reactor module 4 can carry reactants and suspend the reactants in the reactor module 4.

[0084] In some embodiments, the power module 1 includes a DC power supply 6 and a switch 7;

[0085] The control module 2 includes a system controller 8, an input device 9 and an output device 10;

[0086] The liquid processing module 3 includes a valve 11 or a combination of the valves 11, a pump 12 or a combination of the pumps 12 and a sample pool 13;

[0087] The reactor module 4 includes a reactor 14; the reactor 14 includes a reactor frame 15 and a reactor cavity 16 formed by the reactor frame 15;

[0088] The power module 1 is connected to the control module 2, the liquid processing module 3, and the reactor module 4 respectively through cables 25;

[0089] The control module 2 is connected to the liquid processing module 3 and the reactor module 4 respectively through cables 25;

[0090] The liquid treatment module 3 is connected to the reactor module 4 via a pipeline 26;

[0091] The control module 2 is connected to the reactor module 4 via the sensor 5 .

[0092] In some embodiments, the reactor module 4 uses liquid flow or air flow to suspend the reactants in the reactor 14. A liquid film or air film may be formed between the reactants and the wall of the reactor 14 to prevent the reactants from forming a wall adhesion effect.

[0093] In some embodiments, the reactor 14 is one or more and can withstand pressure.

[0094] In some embodiments, when there are multiple reactors 14, they can be connected in series or in parallel.

[0095] In some embodiments, the reactor 14 can be a disposable reactor or a reactor that can be used multiple times.

[0096] In some embodiments, the reactor 14 has at least one liquid inlet and outlet for the entry and exit of reaction liquid.

[0097] In some embodiments, the inner wall of the reactor 14 is smooth or has a certain structure, such as a diverter block 24, which is used to divert the liquid so that the liquid is evenly distributed in the reactor.

[0098] In some embodiments, the inner wall of the reactor 14 may be treated to be hydrophobic or hydrophilic.

[0099] In some embodiments, the hydrophobization may be one or more of silylation and alkylation.

[0100] In some embodiments, the hydrophilization may be one or more of hydroxylation, carboxylation, and amination.

[0101] In some embodiments, the reactor frame 15 is further provided with a pressing component 18 to ensure the sealing performance after the reactor is closed.

[0102] In some embodiments, the material of the reactor 14 is a metal material, such as stainless steel, aluminum alloy, etc.; or a polymer material, such as polypropylene, acrylonitrile-styrene-butadiene copolymer, etc.

[0103] In some embodiments, the reactor frame 15 is further provided with a sealing component 19 , and the material of the sealing component 19 is an elastic material or a material that repels the reaction solution.

[0104] In some embodiments, the reactor module 4 further comprises an identification module, and the bioreactor provided by the present invention feeds back the type of the reactor and the existing procedures through the identification results.

[0105] In some embodiments, RFID is used as the identification module. The reactor module 4 is provided with an identifier, and the reactor is provided with a serial number chip. When the reactor 14 is transferred to the reactor module 4, the identifier identifies the serial number of the chip, and the serial number has a corresponding or no corresponding program in the reactor module 4. For an existing program, the reactor module 4 will call the program. For a program that does not have a corresponding program, the program can be set and saved by the reactor module 4.

[0106] In some embodiments, the reaction is continued after being interrupted by an identification method. For example, after a reaction liquid has reacted, the reactor 14 is taken out from the reactor module 4 to interrupt the process. When the reactor 14 is put into the reactor module 4 again, the reactor module 4 identifies that there is an unfinished program, and displays the unfinished program on the interface, prompting the operator whether to continue the program.

[0107] In some embodiments, a mother liquor pool 23 is further included before the reactor 14 , and the reaction solution is diluted and carried out through the mother liquor pool 23 by reaction dilution to form a final reaction solution in the reactor 14 .

[0108] In some embodiments, the object to be reacted is a biological macromolecule carrier, such as a film, polyacrylamide gel, agarose gel, etc.

[0109] In some embodiments, when the reactant is a film, the reactor 14 forms a micro cavity. When the reactant is a gel, the micro cavity is the reactor chamber 16, which can withstand positive pressure or negative pressure.

[0110] In some embodiments, the liquid processing module 3 comprises one or more sample placement containers, namely, sample pools 13 .

[0111] In some embodiments, the liquid processing module 3 comprises at least one unidirectional and / or bidirectional pump 12 .

[0112] In some embodiments, the pump 12 is one or more of a peristaltic pump, a diaphragm pump, a gear pump, and a plunger pump.

[0113] In some embodiments, the reaction solution is pumped into or out of the reactor 14 by the pump 12 .

[0114] In some embodiments, the liquid processing module 3 includes a valve 11 or a combination of valves 11.

[0115] In some embodiments, the valve 11 is one or more of a diaphragm valve, a pinch valve, and a column valve.

[0116] In some embodiments, the liquid processing module 3 can use two or more different solutions and handle the automatic exchange between these solutions in the reactor 14.

[0117] In some embodiments, the reactor 14 is connected to the liquid treatment module 3 through a pipeline 26 or a one-way valve or a two-way quick connector. The two-way quick connector is used to connect the pipeline and the reactor. Its function is to remove the reactor from the system when the reactor contains liquid (Example 17). After the reactor is disconnected from the pipeline, it is closed to prevent leakage or contamination.

[0118] In some embodiments, the bioreactor uses a pump 12 to circulate the reaction liquid in the reactor 14 .

[0119] In some embodiments, the biomacromolecule is one or more of a nucleic acid, a protein, or a polypeptide.

[0120] In some embodiments, the bioreactor device comprises a liquid sensing module.

[0121] In some embodiments, at least one reactor 14 is included. The number of reactors 14 depends on the number of samples in the reactor 14 and the requirements of the reaction solution. For example, one sample and one solution require one reactor 14; two samples and one solution require one reactor 14. Place the sample in the reactor 14, set the solution quantity, sequence, and time, and the system will operate according to the settings. In this bioreactor, the pump 12 is used to pump the solution into and out of the reactor 14. According to the requirements, a cycle can be formed according to the settings to increase the contact time between the sample and the reaction solution.

[0122] In some embodiments, the contact probability can be increased by setting a pulsed liquid feed.

[0123] Specifically, the structure of the biomacromolecule automated reaction system provided by the present invention is as follows:

[0124] Figure 1 The basic composition of the present invention is shown.

[0125] 1 is a power module, which includes a DC power supply 6 and a switch 7. Its function is to convert AC input into DC output to provide energy required for the operation of other components of the present invention.

[0126] 2 is a control module, which includes a system controller 8, an input device 9, and an output device 10. It works through the energy input by the power module 1, transfers the energy provided by the power module 1 to the liquid processing module 3 and the reactor module 4 through the cable 25, and detects the working status of the liquid processing module 3 and the reactor module 4 through the sensor 5 and controls them.

[0127] 3 is a liquid processing module, which includes a valve 11 or a combination of valves, a pump 12 or a combination of pumps and a sample pool 13. The valve 11 or the combination of valves, the pump 12 or the combination of pumps and the reactor 14 are connected by a pipeline (26). The valve 11 or the combination of valves selects samples according to the signal of the control module 2, and the pump 12 or the combination of pumps pumps the sample into or out of the reactor 14. The sensor 5 is used to detect the operating state of the liquid processing module 3 and feed it back to the control module 2. The operating energy of the liquid processing module 3 is transmitted from the power module 1 by the control module 2, and the power module 1 can also provide energy directly.

[0128] 4 is a reactor module, wherein 14 is a reactor, 17 is an opening provided on the reactor frame 15, which is a liquid inlet 17-1 or a liquid outlet 17-2, and liquid is pumped into or out of the reactor 14 by a pump 12 or a combination of pumps through a pipeline connecting the opening 17. The sensor 5 detects the working state of the reactor 14 and feeds back to the control module 2. The operating capacity of the reactor module 4 is transmitted from the power module 1 by the control module 2, and the capacity can also be directly provided by the power module 1.

[0129] Figure 2 An application example of the present invention is shown, which includes a reactor, four reaction liquids, a pump, and six two-position three-way valves. Among them: 13 is four sample pools, each of which can be filled with reaction liquid. 12 is a pump, specifically a peristaltic pump. 14 is a reactor. 17-1 is a liquid inlet. 17-2 is a liquid outlet. 23 is a mother liquid pool. 27 is a waste liquid collector. 28 is a liquid level schematic component. 29 is a reaction liquid inlet and outlet pipeline. 30 is a waste liquid discharge or air inlet pipeline. 50, 51, 52, 53, 54, 55 are two-position three-way valves. 32, 33, 34 are three liquid receiving ports of a two-position three-way valve, 33 is a common end, and 32 and 33 are normally open. 38, 39, 40 are three liquid receiving ports of a two-position three-way valve, 39 is a common end, and 38 and 39 are normally open. 35, 36, 37 are three liquid receiving ports of a two-position three-way valve, 36 is a common end, and 35 and 36 are normally open. 41, 42, 43 are three liquid receiving ports of a two-position three-way valve, 43 is a common end, and 42 and 43 are normally open. 44, 45, 46 are three liquid receiving ports of a two-position three-way valve, 44 is a common end, and 46 and 44 are normally open. 47, 48, 49 are three liquid receiving ports of a two-position three-way valve, 49 is a common end, and 47 and 49 are normally open. 28 must be maintained below the liquid level of the container to ensure that enough liquid can enter during operation. 30 must be maintained above the liquid level of the container to ensure that waste liquid will not be sucked into the pipeline when air is inhaled. Furthermore, a gas-liquid selection valve can be added at the end of 30 to ensure that air intake and liquid discharge are completely isolated.

[0130] Figure 3 An application example of the present invention is shown, which includes three reactors 14, six reaction liquids, three pumps 12, and two valves 11 or valve groups. Among them: 62 is a gas buffer bottle. 27 is a waste liquid collector. 13 is a sample pool. 11 is a valve, specifically an eight-position selection valve. 66, 67, 68, 69, 70, 71, 72, 73 are selection interfaces of the eight-position valve. 74 is a common interface. 65 is a selection path, which connects the selection interfaces 66, 67, 68, 69, 70, 71, 72, 73 with the common interface 74 according to the signal of the control module 2. 64 is a four-position selection valve group. 12 is a pump, specifically a plunger pump. 14 is a reactor.

[0131] Figure 4 The flow direction of the liquid on the time axis in the reactor 14 is shown. Furthermore, the liquid stationary time can be increased to enhance the effect.

[0132] Figure 5A The schematic diagram of the reactor is shown. 19 is a sealing component, 20 is a front plate of the reactor, 21 is a rear plate of the reactor, 18-1 is a front pressing structure of the reactor, and 18-2 is a rear pressing structure of the reactor. 18-1 and 20 can be integrated, and 21 and 18-2 can be integrated. When the reactor frame 15 of the reactor 14 is closed, a reactor cavity 16 can be formed. The size of the reactor cavity 16 can be determined by the sealing component 19 and the pressing component 18. When the reactant is a film, the reactor cavity 16 at this time forms a micro cavity. Furthermore, a limiting component 22 can be added inside 20 and 21 to maintain a certain cavity. Furthermore, the sealing component 19 can be omitted, and a sealed cavity is directly formed by the tightness of 20 and 21. There is at least one opening 17 on the reactor 14, 17-1 is a liquid inlet; 17-2 is a liquid outlet; the opening 17 is connected to the pump 12 or the valve 11 or the reactor 14 for the input or output of the sample.

[0133] Figure 5B A reactor design was presented. Figure 5C is a cross-sectional view, Figure 5D 18 is a clamping component; 91 is a rotating shaft of the clamping component, through which the clamping component 18 is fixed to the reactor frame 15 and can rotate around it; 20 is a reactor front plate; 21 is a reactor rear plate; 15 is a reactor frame; 17 is an opening, which passes through the reactor frame 15 and is connected to the reactor rear plate 21; 19 is a sealing component; and 16 is a formed reactor cavity.

[0134] Figure 5E A reactor design was presented. Fig. 5F is a cross-sectional view, Figure 5G It is a rear view. Among them: 96 is the reactor cover; 15 is the reactor frame; 20 is the reactor front plate; 21 is the reactor rear plate; 99 is the connecting shaft between the reactor cover 96 and the reactor frame 15; 18 is the pressing component; 19 is the sealing component; 22 is the limiting component; 16 is the reactor cavity; 17 is the opening.

[0135] Fig. 6A , 6B, 6C: wherein 106 is the inner wall of the reactor 14. 17 is the opening. 107 is the reactant. 19 is the sealing component. 109 is a schematic diagram of the liquid flow.

[0136] Fig. 6AThe state of the reactant 107 in the reactor 14 when it is not suspended is shown. The reactant 107 has an irregular shape and may be partially attached to the inner wall 106 of the reactor 14 .

[0137] Figure 6B 6C shows a schematic diagram of suspending the reactant by liquid. When the liquid enters the reactor 14, the liquid flows through the gap between the reactant and the reactor 14, suspending the reactant in the reactor 14. The liquid is fed through the liquid inlets on both sides, and the reactant is suspended in the reactor 14 by the liquid flow. Further, as Figure 6B As shown, the reactant can be suspended by single-side liquid inlet.

[0138] Figure 7 It is shown that the liquid flow is divided by adding an opening 17 or a flow divider 24 to achieve uniform distribution of the liquid entering the reactor 14. Among them: 17 is an opening, specifically a liquid inlet; 19 is a sealing component, specifically a sealing ring; 16 is a reactor cavity; 24 is a flow divider inside the reactor 14.

[0139] Fig. 8A This is a schematic diagram of a set of bidirectional quick plugs, which consists of quick plug A112 and quick plug B113; Figure 8B This is a schematic diagram of the quick plug-in B113; Figure 8C This is a cross-sectional view of the quick plug-in B113; Fig.8D This is a schematic diagram of the quick plug-in A112; Fig. 8E114 is a pagoda joint of the quick-plug A112, which is a derivative of 125 and is used to connect pipelines or directly connect to the container wall; 125 is a closure of the quick-plug B113, which forms a cavity with 116; 118 is the internal flow channel of 114; 115 is the pagoda pattern inside the quick-plug B113, which is used for internal leakage prevention and anti-falling after the quick-plug A and the quick-plug B are assembled; 116 is the main body of the quick-plug B113; 117 is a conducting member; 119 is the internal flow channel of the conducting member; 120 is a guide structure of the quick-plug B113, which is used for internal guiding and fixing after being combined with the quick-plug A; 121 is a limiting structure of the conducting member, which prevents the conducting member 117 from falling off the quick-plug B113; 122 is an internal limiting structure of the quick-plug B113, which limits the inner retreat distance of the conducting member 117; 124 is a quick-plug B113 The outer limit structure limits the outward distance of the conducting member 117; 123 is a structural member with a one-way valve function, which controls the quick plug-in B113 to be closed from the direction of 118 to 119 and communicated from the direction of 119 to 118 when it is separated from the quick plug-in A112; 129 is the upper closing structure of the quick plug-in A112, which is used to fix the leak-proof ring 130; 130 is the leak-proof ring of the quick plug-in A112; 128 is the main body of the quick plug-in A112; 127 is the lower closing structure of the quick plug-in A112; 126 is the pagoda joint of the quick plug-in A112; 131 is the guide structure of the quick plug-in A112; 132 is the internal flow channel of 126; 133 is a structural member with a one-way valve function, which controls the quick plug-in A113 to be closed from the direction of 132 to 129 and communicated from the direction of 129 to 132 when it is separated from the quick plug-in A112. When the quick plug-in A and the quick plug-in B are combined, the conducting member 119 retreats to 122, and 123 of 113 is opened, so that it loses the one-way valve function, and 119 is inserted into 123 of 112, so that it loses the one-way valve function.

[0140] Another aspect of the present invention protects a method for detecting biomacromolecules using the biomacromolecule automated reaction system.

[0141] The beneficial effects of the present invention are: saving time. The system is fully automatic, saving a lot of manual work and greatly improving efficiency. High sensitivity. Through liquid flow control, the reaction efficiency is improved compared with the traditional method, and the result sensitivity is higher under the same conditions as the traditional method. Saving reactor reagents. In this system, the reactor only needs a small amount of liquid to complete the reaction of a large amount of reagents in the traditional method.

[0142] The raw materials and reagents used in the bioreactor provided by the present invention and the method for performing bioassay based on the bioreactor can all be purchased from the market.

[0143] The present invention will be further described below in conjunction with embodiments:

[0144] Example 1 Bioreactor provided by the present invention

[0145] The present invention provides a bioreactor, such as Figure 1 As shown, it includes a power module 1, a control module 2, a liquid processing module 3, a reactor module 4 and a sensor 5;

[0146] in:

[0147] 1 is a power module, which includes a DC power supply 6 and a switch 7. Its function is to convert AC input into DC output to provide energy required for the operation of other components of the present invention.

[0148] 2 is a control module, which includes a system controller 8, an input device 9, and an output device 10. It works through the energy input by the power module 1, transfers the energy provided by the power module 1 to the liquid processing module 3 and the reactor module 4 through the cable (25), and detects the working status of the liquid processing module 3 and the reactor module 4 through the sensor 5 and controls them.

[0149] 3 is a liquid processing module, which includes a valve 11 or a combination of valves, a pump 12 or a combination of pumps and a sample pool 13. The valve 11 or the combination of valves, the pump 12 or the combination of pumps and the reactor 14 are connected by a pipeline (26). The valve 11 or the combination of valves selects samples according to the signal of the control module 2, and the pump 12 or the combination of pumps pumps the sample into or out of the reactor 14. The sensor 5 is used to detect the operating state of the liquid processing module 3 and feed it back to the control module 2. The operating energy of the liquid processing module 3 is transmitted from the power module 1 by the control module 2, and the power module 1 can also provide energy directly.

[0150] 4 is a reactor module, wherein 14 is a reactor, including a reactor frame 15 and a reactor cavity 16 formed by the reactor frame 15; 17 is an opening provided on the reactor frame 15, which is a liquid inlet 17-1 or a liquid outlet 17-2, and liquid is pumped into or out of the reactor 14 by a pump 12 or a combination of pumps through a pipeline connecting the opening 17. The sensor 5 detects the working state of the reactor 14 and feeds back to the control module 2. The operating capacity of the reactor module 4 is transmitted from the power module 1 by the control module 2, and the capacity can also be directly provided by the power module 1.

[0151] The power module 1 is connected to the control module 2, the liquid processing module 3, and the reactor module 4 through cables 25; the control module 2 is connected to the liquid processing module 3 and the reactor module 4 through cables 25; the liquid processing module 3 is connected to the reactor module 4 through a pipeline 26; the control module 2 is connected to the reactor module 4 through a sensor 5, and the opening 17 is connected to the pump 12 or a combination of pumps 12.

[0152] Example 2

[0153] Based on the structure of the bioreactor provided in Example 1, the present invention further provides a bioreactor such as Figure 2 As shown, it includes a reactor, four reaction liquids, a pump, and 6 two-position three-way valves.

[0154] Among them: 13 is four sample pools, each sample pool 13 can be filled with reaction liquid. 12 is a pump, specifically a peristaltic pump. 14 is a reactor. 17-1 is a liquid inlet. 17-2 is a liquid outlet. 23 is a mother liquid pool. 27 is a waste liquid collector. 28 is a liquid level schematic component. 29 is a reaction liquid inlet and outlet pipeline. 30 is a waste liquid discharge or air inlet pipeline. 50, 51, 52, 53, 54, 55 are two-position three-way valves. 32, 33, 34 are three liquid receiving ports of a two-position three-way valve, 33 is a common end, and 32 and 33 are normally open. 38, 39, 40 are three liquid receiving ports of a two-position three-way valve, 39 is a common end, and 38 and 39 are normally open. 35, 36, 37 are three liquid receiving ports of a two-position three-way valve, 36 is a common end, and 35 and 36 are normally open. 41, 42, 43 are three liquid connection ports of a two-position three-way valve, 43 is a common end, and 42 and 43 are normally open. 44, 45, 46 are three liquid connection ports of a two-position three-way valve, 44 is a common end, and 46 and 44 are normally open. 47, 48, 49 are three liquid connection ports of a two-position three-way valve, 49 is a common end, and 47 and 49 are normally open.

[0155] 28 must be maintained below the liquid level in the container to ensure that sufficient liquid can enter during operation. 30 must be maintained above the liquid level in the container to ensure that waste liquid is not sucked into the pipeline during air intake. Furthermore, a gas-liquid selection valve can be added at the end of 30 to ensure that air intake and liquid discharge are completely isolated.

[0156] Example 3

[0157] Based on the structure of the bioreactor provided in Example 1, the present invention further provides a bioreactor such as Figure 3 As shown, three reactors 14, six reaction liquids, three pumps 12, and two valves 11 or valve groups are included.

[0158] Among them: 62 is a gas buffer bottle. 27 is a waste liquid collector. 13 is a sample pool. 11 is a valve, specifically an eight-position selection valve. 66, 67, 68, 69, 70, 71, 72, 73 are selection interfaces of the eight-position valve. 74 is a common interface. 65 is a selection path, which connects the selection interfaces 66, 67, 68, 69, 70, 71, 72, 73 with the common interface 74 according to the signal of the control module 2. 64 is a four-position selection valve group. 12 is a pump, specifically a plunger pump. 14 is a reactor. The reactors can be connected in series or in parallel. For example, the reactor 14-1 and the reactor 14-2 are in series.

[0159] Example 4

[0160] Based on the structure of the bioreactor provided in Example 1, the present invention further provides a bioreactor, as shown in FIG5(A), wherein 19 is a sealing component, 20 is a reactor front plate, 21 is a reactor rear plate, 18-1 is a reactor front pressing structure, and 18-2 is a reactor rear pressing structure. 18-1 and 20 can be integrated, and 21 and 18-2 can be integrated. When the reactor frame 15 of the reactor 14 is closed, a reactor cavity 16 can be formed. The size of the reactor cavity 16 can be determined by the sealing component 19 and the pressing component 18. When the reactant is a film, the reactor cavity 16 at this time forms a micro cavity. Furthermore, a limiting component 22 can be added inside 20 and 21 to maintain a certain cavity. Furthermore, the sealing component 19 can be omitted, and the sealed cavity is directly formed by the tightness of 20 and 21. There is at least one opening 17 on the reactor 14, 17-1 is the liquid inlet; 17-2 is the liquid outlet; the opening 17 is connected to the pump 12 or the valve 11 or the reactor 14 for inputting or outputting samples.

[0161] Example 5

[0162] Based on the structure of the bioreactor provided in Example 1, the present invention further provides a bioreactor such as Figure 5(B) to Figure 5(D) As shown, wherein: 18 is a clamping component; 91 is a rotating shaft of the clamping component, through which the clamping component 18 is fixed to the reactor frame 15 and can rotate around it; 20 is a reactor front plate; 21 is a reactor rear plate; 15 is a reactor frame; 17 is an opening, which passes through the reactor frame 15 and is connected to the reactor rear plate 21; 19 is a sealing component; and 16 is a formed reactor cavity.

[0163] Example 6

[0164] Based on the structure of the bioreactor provided in Example 1, the present invention further provides a bioreactor such as Figure 5(E) to Figure 5(G) As shown, wherein: 96 is the reactor cover; 15 is the reactor frame; 20 is the reactor front plate; 21 is the reactor rear plate; 99 is the connecting shaft between the cover and the frame; 18 is the pressing component; 19 is the sealing component; 22 is the limiting component; 16 is the reactor cavity; and 17 is the opening.

[0165] Example 7

[0166] Based on the structure of the bioreactor provided in Example 1, the present invention further provides a bioreactor, as shown in FIG6 . 106 is the inner wall of the reactor 14 . 17 is the opening . 107 is the reactant . 19 is the sealing component . 109 is a schematic diagram of the liquid flow .

[0167] FIG6(A) shows the state of the reactant 107 in the reactor 14 when it is not suspended. The reactant 107 is in an irregular shape and may be partially attached to the inner wall 106 of the reactor 14 .

[0168] FIG. 6(B) and FIG. 6(C) show schematic diagrams of suspending the reactants by liquid.

[0169] When the liquid enters the reactor 14, the liquid flows through the gap between the reactant and the reactor 14, and the reactant is suspended in the reactor 14. The liquid is fed through the liquid inlets on both sides, and the reactant is suspended in the reactor 14 by the liquid flow. Figure 6B As shown, the reactant can be suspended by single-side liquid inlet.

[0170] Example 8

[0171] Based on the structure of the bioreactor provided in Example 1, the present invention further provides a bioreactor such as Figure 7 As shown, 17 is an opening, specifically a liquid inlet; 19 is a sealing component, specifically a sealing ring; 16 is a reactor cavity; and 24 is a flow divider inside the reactor 14.

[0172] Example 9

[0173] Based on the structure of the bioreactor provided in Example 1, the present invention further provides a bioreactor, as shown in FIG8 . Fig. 8A This is a schematic diagram of a set of bidirectional quick plugs, which consists of quick plug A112 and quick plug B113; Figure 8B This is a schematic diagram of the quick plug-in B113; Figure 8C This is a cross-sectional view of the quick plug-in B113; Fig.8D This is a schematic diagram of the quick plug-in A112; Fig. 8E This is the cross-sectional view of the quick plug-in A112;

[0174] Among them, 114 is the pagoda joint of the quick plug-in B113, which is a derivative part of 125 and is used to connect the pipeline or directly connect to the container wall; 125 is the closing part of the quick plug-in B113, which forms a cavity with 116; 118 is the internal flow channel of 114; 115 is the internal pagoda pattern of the quick plug-in B113, which is used for internal leakage prevention and anti-falling after the quick plug-in A and the quick plug-in B are assembled; 116 is the main body of the quick plug-in B113; 117 is the conductive part; 119 is the internal flow channel of the conductive part; 120 is the guide structure of the quick plug-in B113, which is used for internal guiding and fixing after being combined with the quick plug-in A; 121 is the limiting structure of the conductive part, which prevents the conductive part 117 from falling off from the quick plug-in B113; 122 is the internal limiting structure of the quick plug-in B113, which limits the inner retreat distance of the conductive part 117; 124 is the external limiting structure of the quick plug-in B113 , limiting the outward distance of the conducting member 117; 123 is a structural member with a one-way valve function, which controls the quick plug-in B113 to be closed from the direction of 118 to 119 and communicated from the direction of 119 to 118 when it is separated from the quick plug-in A112; 129 is the upper closing structure of the quick plug-in A112, which is used to fix the leak-proof ring 130; 130 is the leak-proof ring of the quick plug-in A112; 128 is the main body of the quick plug-in A112; 127 is the lower closing structure of the quick plug-in A112; 126 is the pagoda joint of the quick plug-in A112; 131 is the guide structure of the quick plug-in A112; 132 is the internal flow channel of 126; 133 is a structural member with a one-way valve function, which controls the quick plug-in A113 to be closed from the direction of 132 to 129 and communicated from the direction of 129 to 132 when it is separated from the quick plug-in A112. When the quick plug-in A and the quick plug-in B are combined, the conducting member 119 retreats to 122, and 123 of 113 is opened, so that it loses the one-way valve function, and 119 is inserted into 123 of 112, so that it loses the one-way valve function.

[0175] Example 10 Coomassie Brilliant Blue Staining

[0176] Based on the bioreactor provided in Example 1, the staining steps of the present invention are as follows:

[0177] 1. Add the corresponding working solution into the staining solution and decolorizing solution storage bottles;

[0178] 2. Place the gel or membrane to be stained into the reactor;

[0179] 3. Set the instrument working procedure:

[0180] The temperature is 20-30°C, the area to be dyed is 7.5*8CM, and the flow rate is set to 40ml / min.

[0181] A. Dyeing solution inlet (100ml)

[0182] B. The dye solution circulates for 1 hour

[0183] C. Dyeing liquid drainage

[0184] D. Decolorizing solution inlet (100ml)

[0185] E. Decolorization solution circulation for 1h

[0186] F. Drainage of decolorizing solution

[0187] G. Decolorizing solution inlet (100ml)

[0188] H. Decolorization solution circulation for 1h

[0189] I. Drainage of decolorizing solution

[0190] J. End of instrument work

[0191] 4. Take out the gel and record the results

[0192] Traditional dyeing process

[0193] 1. Place the gel or membrane in the box

[0194] 2. Pour 200ml of dye solution

[0195] 3. Oscillate at room temperature for 1 hour

[0196] 4. Discard the staining solution and pour in 200ml of decolorizing solution

[0197] 5. Oscillate at room temperature for 1 hour

[0198] 6. Discard the decolorizing solution in step 4 and pour in 200 ml of decolorizing solution

[0199] 7. 1h at room temperature

[0200] 8. 6-7, until the stripes are clear and the background is clean

[0201] The experimental samples are

[0202] Protein marker (genscript M00521), bovine serum albumin (BSA), E. coli cell lysis, mouse IgG, lysozyme

[0203] The loading volume is:

[0204] 1: marker 5μL

[0205] 2: E. coli cell lysis 20μL

[0206] 3: e.coli cell lysis 16μL

[0207] 4: e.coli cell lysis 12μL

[0208] 5: e.coli cell lysis 8μL

[0209] 6: e.coli cell lysis 4μL

[0210] 7:maker 5μL

[0211] 8: 1000ng IgG, 1000ng BSA, 1000ng lysozyme

[0212] 9: 500ng IgG, 500ng BSA, 500ng lysozyme

[0213] 10: 250ng IgG, 250ng BSA, 250ng lysozyme

[0214] 11: 125ng IgG, 125ng BSA, 125ng lysozyme

[0215] 12: 62.5ng IgG, 62.5ng BSA, 62.5ng lysozyme

[0216] Staining formula: Staining solution: 0.1% R250, 30% ethanol, 10% acetic acid, 60% water; Decolorizing solution: 10% ethanol, 10% acetic acid, 80% water.

[0217] Figure 9 (A) is the staining result of the traditional method, Figure 9 (B) is the staining result of the present invention, Figure 9 (C) shows the comparative analysis of IgG coloring degree in the staining results of the traditional method and the staining results of the present invention; Figure 9 (D) shows the comparative analysis of lysozyme coloring degree in the staining results of the traditional method and the staining results of the present invention; Figure 9 (E) shows the comparative analysis of BSA coloring degree in the staining results of the traditional method and the staining results of the present invention; the results are presented on the same plane using a bio-rad photosensitivity scanner.

[0218] In the traditional dyeing method, a piece of gel needs at least 600 ml of liquid to react, but in the present invention, only 300 ml of liquid is needed to complete the reaction.

[0219] Result analysis:

[0220] Table 1

[0221]

[0222] Table 2

[0223] IgG Traditional methods The present invention Compared with the traditional method, the present invention improves Lane 8 60.7 74.6 22.90％ Lane 9 48.5 51.2 5.57％ Lane 10 37.9 39.6 4.49％ Lane 11 29.1 33.7 15.81％ Lane 12 29.5 31.6 7.12％

[0224] Table 3

[0225] BSA Traditional methods The present invention Compared with the traditional method, the present invention improves Lane 8 45.7 65.9 44.20％ Lane 9 41.7 47.9 14.87％ Lane 10 31.5 39.2 24.44％ Lane 11 22.7 33.7 48.46％ Lane 12 20 32 60.00％

[0226] Table 4

[0227] Lysozyme Traditional methods The present invention Compared with the traditional method, the present invention improves Lane 8 37.1 41 10.51％ Lane 9 28.8 31.6 9.72％ Lane 10 20.8 26.7 28.37％ Lane 11 22.2 24.4 9.91％ Lane 12 0 0 ——

[0228] On IgG, all bands were visible in both the traditional method and the present invention, but the present invention was more deeply stained.

[0229] On BSA, the traditional method can see the 10th sample, while the present invention can see all 12 samples, and the present invention has better sensitivity.

[0230] For lysozyme, the traditional method can detect the 9th sample, while the present invention can detect the 11th sample, and the present invention has better sensitivity.

[0231] As shown in Figures 9(C), 9(D), 9(E) and Tables 2 to 4, compared with the traditional method, the coloring degree of the IgG test results of the present invention is improved by 4.49% to 22.90%; the coloring degree of the BSA test results is improved by 14.87% to 60.00%; and the coloring degree of the lysozyme test results is improved by 9.72% to 28.37%.

[0232] In summary, compared with the traditional method, the present invention only needs to use 50% of the reagents, and the sensitivity and coloring depth are better than the traditional method.

[0233] Example 11 Silver staining

[0234] Based on the bioreactor provided in Example 2, the reaction is as follows:

[0235] Fixative: 40% methanol and 100ul formaldehyde per 200ml

[0236] Sensitizer: 0.2g / L sodium thiosulfate

[0237] Staining agent: 20% AgNO3 250ul / 50ml H2O

[0238] Color development solution: 3% sodium carbonate and 0.0004% sodium thiosulfate. Add 25ul formaldehyde solution / 50ml before use.

[0239] The experimental samples are:

[0240] Protein marker (genscript M00521), E. coli cell lysis, mouse IgG, bovine serum albumin (BSA)

[0241] The loading volume is:

[0242] 1: marker 5μL

[0243] 2: E. coli cell lysis 10μL

[0244] 3: e.coli cell lysis 8μL

[0245] 4: e.coli cell lysis 6μL

[0246] 5: e.coli cell lysis 4μL

[0247] 6: e.coli cell lysis 1μL

[0248] 7:maker 5μL

[0249] 8: 100ng IgG, 100ng BSA

[0250] 9: 50ng IgG, 50ng BSA

[0251] 10: 25ng IgG, 25ng BSA

[0252] 11: 12.5ng IgG, 12.5ng BSA

[0253] 12: 6.25ng IgG, 6.25ng BSA

[0254] 13: 3.125ng IgG, 3.125ng BSA

[0255] Steps of the present invention:

[0256] 1. Place the gel into the reactor

[0257] 2. The setting procedure is as follows

[0258] The temperature is 20-30°C, and the gel area is 7.5*8CM. Compared with Case 10, the reaction rate of the silver staining reagent is lower than that of Coomassie Brilliant Blue, and the flow rate needs to be reduced. The flow rate is set to 20ml / min.

[0259] 1) Pump in 100 ml of fixative solution, set the cycle to 10 minutes, and pump out when the reaction is complete.

[0260] 2) Pump in 100 ml of ddH2O, circulate for 5 minutes, and pump out, twice in total

[0261] 3) Pump in 100 ml of sensitizer, circulate for 1 min, and pump out

[0262] 4) Pump in 100 ml of ddH2O, circulate for 1 min, and pump out, twice in total

[0263] 5) Pump in 100 ml of dye, circulate for 10 minutes, and pump out

[0264] 6) Pump in 100 ml of ddH2O, circulate for 1 min, and pump out, twice in total

[0265] 7) Pump in 100 ml of color developing solution, circulate for 10 minutes, and pump out

[0266] 8) Pump in 100 ml of ddH2O, circulate for 5 minutes, and pump out

[0267] 3. Take out the gel and record the results

[0268] Traditional method steps:

[0269] 1. Put the gel into the container

[0270] 2. Pour in 200 ml of fixative solution, shake at room temperature for 10 minutes, and discard the waste liquid.

[0271] 3. Pour in 200ml of ddH2O, shake at room temperature for 5 minutes, and discard the waste liquid.

[0272] 4. Repeat step 3

[0273] 5. Pour in 200 ml of sensitizer, shake at room temperature for 1 min, and discard the waste liquid

[0274] 6. Pour in 200ml of ddH2O, shake at room temperature for 1min, and discard the waste liquid

[0275] 7. Repeat step 6

[0276] 8. Pour in 200ml of dye, shake at room temperature for 10 minutes, and discard the waste liquid

[0277] 9. Pour in 200ml of ddH2O, shake at room temperature for 1min, and discard the waste liquid

[0278] 10. Repeat steps

[0279] 11. Pour 200 ml of color developing solution, shake at room temperature for 10 minutes, and discard the waste liquid

[0280] 12. Pour in 200ml of ddH2O, shake at room temperature for 5 minutes, and discard the waste liquid.

[0281] 13. Take out the gel and record the results

[0282] Figure 10(A) is the staining result of the present invention, and Figure 10(B) is the staining result of the traditional method; Figure 10(C) shows a comparative analysis of the IgG coloring degree in the staining results of the traditional method and the staining results of the present invention; Figure 10(D) shows a comparative analysis of the BSA coloring degree in the staining results of the traditional method and the staining results of the present invention.

[0283] Table 5

[0284]

[0285]

[0286] Table 6

[0287] IgG Traditional methods The present invention Compared with the traditional method, the present invention improves Lane 8 81.8 175.8 114.91％ Lane 9 79.5 177.3 123.02％ Lane 10 76.5 175 128.76％ Lane 11 74 169.6 129.19％ Lane 12 71.3 170.6 139.27％

[0288] Table 7

[0289] BSA Traditional methods The present invention Compared with the traditional method, the present invention improves Lane 8 79.3 159.3 100.88％ Lane 9 70.5 157.3 123.12％ Lane 10 63.7 156.8 146.15％ Lane 11 58 155.2 167.59％ Lane 12 54.7 156.7 186.47％

[0290] As can be seen from the figure, when using half the reagent, the sensitivity of the present invention is better than that of the traditional method, and the background is better than that of the traditional method. On IgG, all bands of the traditional method and the present invention are visible. On BSA, the traditional method can see the 10th sample, while the present invention can see all 12 samples, and the present invention has better sensitivity.

[0291] As shown in Figure 10 (C), Figure 10 (D) and the results of Tables 6 and 7, compared with the traditional method, the coloring degree of IgG detection results of the method provided by the present invention is improved by 114.91% to 139.27%; the coloring degree of BSA detection results is improved by 100.88% to 186.47%.

[0292] In summary, compared with the traditional method, the present invention only needs to use 50% of the reagents, and its sensitivity and coloring degree are better than the traditional method.

[0293] Example 12 WB1

[0294] Based on the bioreactor provided in Example 3, the reaction is as follows:

[0295] The samples are His-tagged proteins with molecular weights of 93kd and 7kd

[0296] The loading amount is

[0297] 1. WB Marker 1ul

[0298] 2. 150ng 93kd, 10ng 7kd

[0299] 3. 75ng 93kd, 5ng 7kd

[0300] 4. 37.5ng 93kd, 2.5ng 7kd

[0301] 5. 18.75ng 93kd, 1.25ng 7kd

[0302] 6. 9ng 93kd, 0.6ng 7kd

[0303] 7, 4.5ng 93kd, 0.3ng 7kd

[0304] 8. 2.25ng 93kd, 0.15ng 7kd

[0305] 9. 1ng 93kd, 0.075ng 7kd

[0306] 10. 0.5ng 93kd, 0.0375ng 7kd

[0307] The blocking solution is 5% skim milk powder in PBS

[0308] PBST is PBS with 1 / 1000 Tween 20

[0309] The primary antibody was mouse anti-His at a concentration of 1 mg / ml, using blocking solution at a dilution of 1:3000

[0310] The secondary antibody was goat anti-mouse HRP at a concentration of 1 mg / ml, using blocking solution at a dilution of 1:10000

[0311] Steps of the present invention:

[0312] 1. Place two membranes back to back in the reactor, the temperature is 20-30°C, the membrane area is 7.5*8CM*2, and the flow rate is set to 40ml / min;

[0313] 2. Pump in 8 ml of blocking solution, circulate for 1 hour, and pump out;

[0314] 3. Pump in 8 ml of PBST, circulate for 5 min, and pump out for 3 times;

[0315] 4. Pump in 8 ml of primary antibody dilution, circulate for 1 hour, and pump out;

[0316] 5. Pump in 8 ml of PBST, circulate for 5 min, and pump out for 3 times;

[0317] 6. Pump in 8 ml of secondary antibody dilution solution, circulate for 1 hour, and pump out;

[0318] 7. Pump in 8 ml of PBST, circulate for 5 min, and pump out for 3 times;

[0319] 8. Take out the film for exposure.

[0320] Traditional method:

[0321] 1. Place two films back to back in a container

[0322] 2. Add 10 ml of blocking solution, shake for 1 hour, and discard the blocking solution.

[0323] 3. Add 10 ml of PBST, shake for 5 minutes, and discard PBST

[0324] 4. Repeat step 3 twice

[0325] 5. Add 10 ml of primary antibody diluent, shake for 1 hour, and discard the primary antibody diluent

[0326] 6. Add 10 ml of PBST, shake for 5 minutes, and discard PBST

[0327] 7. Repeat step 3 three times

[0328] 8. Add 10 ml of secondary antibody diluent, shake for 1 hour, and discard the secondary antibody diluent

[0329] 9. Repeat step 3 three times

[0330] 10. Take out the film for exposure

[0331] Figure 11(A) is the result of the traditional method, and Figure 11(B) is the result of the method provided by the present invention; Figures 11(C) to 11(D) show the results of the method provided by the present invention; Figure 11(E) shows a comparative analysis of the coloring degree of 93kd protein in the results of the traditional method and the results of the present invention; Figure 11(F) shows a comparative analysis of the coloring degree of 7kd protein in the results of the traditional method and the results of the present invention.

[0332] Table 8

[0333]

[0334] Table 9

[0335]

[0336]

[0337] Table 10

[0338]

[0339] As can be seen from FIG. 11 , at 93 kd, the conventional method can be seen in the 4th lane, while the present invention can be seen in the 5th lane, and the present invention has better sensitivity.

[0340] At 7 kd, it can be seen in the 8th lane of the traditional method and in the 9th lane of the present invention, and the sensitivity of the present invention is better.

[0341] As shown in Figure 11 (E), Figure 11 (F) and Tables 9 and 10, compared with the traditional method, the coloring degree of the 93kd detection result provided by the method provided by the present invention is improved by 102.09% to 507.93%; the coloring degree of the 7kd detection result is improved by 20.61% to 432.91%.

[0342] In summary, compared with the traditional method, the present invention only needs to use 74% of the reagents and 80% of the antibodies, and the sensitivity and signal strength of the present invention are better than those of the traditional method.

[0343] Example 13 WB2

[0344] Based on the bioreactor provided in Example 4, the reaction is as follows:

[0345] The sample is 293T cell lysate

[0346] The loading amount is

[0347] 1. WB marker 1ul

[0348] 2. Lysis solution 20ug

[0349] 3. Lysis solution 10ug

[0350] 4. Lysis solution 5ug

[0351] 5. Lysis solution 2.5ug

[0352] 6. Lysis solution 1.25ug

[0353] 7. Lysis solution 0.625ug

[0354] 8. Lysis solution 0.3125ug

[0355] The blocking solution is 5% skim milk powder in PBS

[0356] PBST is PBS with 1 / 1000 Tween 20

[0357] The primary antibody was rabbit anti-β-actin at a concentration of 1 mg / ml, using blocking solution at a dilution of 1:3000

[0358] The secondary antibody was goat anti-rabbit HRP at a concentration of 1 mg / ml, and the blocking solution was used at a dilution of 1:10000.

[0359] Steps of the present invention

[0360] The temperature is 20-30°C, the membrane area is 7.5*8CM, and the flow rate is set to 20ml / min;

[0361] 1. Place a membrane into the reactor

[0362] 2. Pump in the blocking solution

[0363] 3. When the blocking solution fills the reactor, pump in PBST

[0364] 4. When PBST fills the reactor, pump in the primary antibody dilution

[0365] 5. When the primary antibody dilution solution fills the reactor, pump in PBST

[0366] 6. When PBST fills the reactor, pump in the secondary antibody dilution solution

[0367] 7. When the secondary antibody dilution solution fills the reactor, pump in PBST

[0368] 8. When the PBST fills the reactor, continue to pump in 2 times the volume of PBST.

[0369] 9. Remove the film for development

[0370] The blocking solution, primary antibody diluent, and secondary antibody diluent are all 4 ml, and PBST is 16 ml

[0371] It takes a total of 2 hours to complete the use of the present invention

[0372] Traditional method:

[0373] 1. Place the membrane into a container;

[0374] 2. Add 10 ml of blocking solution, shake for 1 hour, and discard the blocking solution;

[0375] 3. Add 10 ml of PBST, shake for 5 minutes, and discard the PBST;

[0376] 4. Repeat step 3 twice;

[0377] 5. Add 10 ml of primary antibody diluent, shake for 1 hour, and discard the primary antibody diluent;

[0378] 6. Add 10 ml of PBST, shake for 5 minutes, and discard the PBST;

[0379] 7. Repeat step 3 three times;

[0380] 8. Add 10 ml of secondary antibody diluent, shake for 1 hour, and discard the secondary antibody diluent;

[0381] 9. Repeat step 3 three times;

[0382] 10. Take out the film for exposure.

[0383] FIG. 12(A) shows the traditional method, FIG. 12(B) shows the result of the present invention; FIG. 12(C) shows the comparative analysis of the β-actin protein staining degree in the traditional method result and the result of the present invention.

[0384] Table 11

[0385]

[0386] Table 12

[0387] β-actin Traditional methods The present invention Compared with the traditional method, the present invention improves Lane 2 109.9 161.7 47.13％ Lane 3 128 131.9 3.05％ Lane 4 111.1 120.1 8.10％ Lane 5 54.7 100.3 83.36％ Lane 6 32.6 73.5 125.46％ Lane 7 7.5 30.4 305.33％ Lane 8 5.2 11.1 113.46％

[0388] As can be seen from the attached drawings, the conventional method can be seen in lane 6, while the present invention can be seen in lane 8, and the present invention has better sensitivity.

[0389] As shown in FIG. 12 (C) and Table 12, compared with the traditional method, the color degree of the β-actin detection result of the method provided by the present invention is improved by 3.05% to 305.33%.

[0390] In summary, compared with the traditional method, the present invention only needs to use 65% of the reagents, 40% of the antibodies and a shorter processing time, and the sensitivity and signal strength of the present invention are better than the traditional method.

[0391] Example 14 WB5

[0392] Based on the bioreactor provided in Example 3, the reaction is as follows:

[0393] The sample is 293T cell lysate

[0394] The loading amount is

[0395] 1. WB marker 1ul

[0396] 2. Lysis solution 20ug

[0397] 3. Lysis solution 10ug

[0398] 4. Lysis solution 5ug

[0399] 5. Lysis solution 2.5ug

[0400] 6. Lysis solution 1.25ug

[0401] The primary antibody was mouse anti-GAPDH

[0402] Secondary antibody: goat anti-mouse

[0403] Steps of the present invention

[0404] 1. Place membrane B into reactor 14-1 and membrane D into reactor 14-2;

[0405] 2. Set reactor 14-1 and reactor 14-2 in series

[0406] The temperature is 20-30°C, the area of ​​membrane B and membrane D is 7.5*4CM, and the flow rate is set to 10ml / min

[0407] 3. Pump in the sealing liquid to fill the reactor 14-1 and the reactor 14-2, circulate for 1 hour, and pump out

[0408] 4. Pump in PBST, circulate for 5 minutes, and pump out for 3 times

[0409] 5. Pump in the primary antibody dilution, circulate for 1 hour, and pump out

[0410] 6. Pump in PBST, circulate for 5 minutes, and pump out for 3 times

[0411] 7. Pump in the secondary antibody dilution solution, circulate for 1 hour, and pump out

[0412] 8. Pump in PBST, circulate for 5 minutes, and pump out for 3 times

[0413] 9. Take out the film for exposure

[0414] The blocking solution, primary antibody diluent, and secondary antibody diluent are all 8 ml, and PBST is 7 ml

[0415] Traditional method:

[0416] 1. Place two films (A, C) in containers respectively.

[0417] 2. Add 10 ml of blocking solution to each tube, shake for 1 hour, and discard the blocking solution.

[0418] 3. Add 10 ml of PBST to each tube, shake for 5 minutes, and discard PBST.

[0419] 4. Repeat step 3 twice

[0420] 5. Add 10 ml of primary antibody diluent to each tube, shake for 1 hour, and discard the primary antibody diluent.

[0421] 6. Add 10 ml of PBST to each tube, shake for 5 minutes, and discard PBST.

[0422] 7. Repeat step 3 three times

[0423] 8. Add 10 ml of secondary antibody diluent to each tube, shake for 1 hour, and discard the secondary antibody diluent.

[0424] 9. Repeat step 3 three times

[0425] 10. Take out the film for exposure

[0426] Figure 13 (A) is the result of membrane A, Figure 13 (B) is the result of membrane B, Figure 13 (C) is the result of membrane C, and Figure 13 (D) is the result of membrane D; Figure 13 (E) shows a comparative analysis of the coloring degree of GAPDH protein in the results of the traditional method and the results of the present invention.

[0427] Table 13

[0428] GAPDH Lane 2 Lane 3 Lane 4 Lane 5 Membrane A 79.7 60.3 29.9 9.4 Membrane B 154.3 124.4 43.1 12.3 Membrane C 84.5 54 20.6 3.9 Membrane D 143.6 114.3 44.6 7.7 The average value of film A and film C 82.1 57.15 25.25 6.65 The average value of membrane B and membrane D 148.95 119.35 43.85 10 Compared with the traditional method, the present invention improves 81.43％ 108.84％ 73.66％ 50.38％

[0429] As can be seen from the figure, the colors of Figure 13 (B) and Figure 13 (D) are darker than those of Figure 13 (A) and Figure 13 (C). As can be seen from Figure 13 (E) and Table 13, the coloring degree of the GAPDH detection result of the method provided by the present invention is increased by 50.38% to 108.84% compared with the traditional method. That is, its signal intensity is greater, further indicating that the effect of the present invention is better than that of the traditional method. In summary, compared with the traditional method, the present invention only needs to use 66% of the reagents, and the sensitivity and signal intensity of the present invention are better than those of the traditional method.

[0430] Example 15 WB3

[0431] Based on the bioreactor provided in Example 5, the reaction is as follows:

[0432] The sample is 293T cell lysate

[0433] The loading amount is

[0434] 1. Lysis solution 20ug

[0435] 2. Lysis solution 10ug

[0436] 3. Lysis solution 5ug

[0437] 4. Lysis solution 2.5ug

[0438] 5. Lysis solution 1.25ug

[0439] 6. Lysis solution 0.625ug

[0440] The blocking solution is 5% skim milk powder in PBS

[0441] PBST is PBS with 1 / 1000 Tween 20

[0442] The primary antibody was rabbit anti-β-actin at a concentration of 1 mg / ml, using blocking solution at a dilution of 1:3000

[0443] The secondary antibody was goat anti-rabbit HRP at a concentration of 1 mg / ml, and the blocking solution was used at a dilution of 1:10000.

[0444] Steps of the present invention:

[0445] 1. The temperature is 20-30°C, the membrane area is 7.5*4CM, and the flow rate is set to 10ml / min;

[0446] 2. Pump in 5 ml of blocking solution, circulate for 1 hour, and pump out;

[0447] 3. Pump in 5 ml of PBST, circulate for 5 min, and pump out for 3 times;

[0448] 4. Pump in 5 ml of primary antibody and secondary antibody mixed solution, circulate for 1 hour, and pump out for recovery;

[0449] 5. Pump in 5 ml of PBST, circulate for 5 min, and pump out for 3 times;

[0450] 6. Pump in the developer and pump out when it is full;

[0451] 7. Take out the film for development;

[0452] Traditional method:

[0453] 1. Place two films back to back in a container;

[0454] 2. Add 10 ml of blocking solution, shake for 1 hour, and discard the blocking solution;

[0455] 3. Add 10 ml of PBST, shake for 5 minutes, and discard the PBST;

[0456] 4. Repeat step 3 twice;

[0457] 5. Add 10 ml of primary antibody diluent, shake for 1 hour, and discard the primary antibody diluent;

[0458] 6. Add 10 ml of PBST, shake for 5 minutes, and discard the PBST;

[0459] 7. Repeat step 3 three times;

[0460] 8. Add 10 ml of secondary antibody diluent, shake for 1 hour, and discard the secondary antibody diluent;

[0461] 9. Repeat step 3 three times;

[0462] 10. Take out the film for exposure.

[0463] FIG. 14(A) shows the result of the traditional method, and FIG. 14(B) shows the result of the present invention; FIG. 14(C) shows a comparative analysis of the staining degree of β-actin protein in the result of the traditional method and the result of the present invention.

[0464] Table 14

[0465]

[0466]

[0467] Table 15

[0468] β-actin Traditional methods The present invention Compared with the traditional method, the present invention improves Lane 1 171.9 219.2 27.52％ Lane 2 122.8 212 72.64％ Lane 3 82.7 183.2 121.52％ Lane 4 16.7 79.7 377.25％ Lane 5 2.8 19.7 603.57％ Lane 6 2.9 4.7 62.07％

[0469] As can be seen from FIG. 14 , the conventional method can be seen in lane 4, while the present invention can be seen in lane 5, and the present invention has better sensitivity.

[0470] As shown in FIG. 14(C) and Table 15, compared with the traditional method, the coloring degree of β-actin detection results of the method provided by the present invention is improved by 27.52% to 603.57%.

[0471] In summary, compared with the traditional method, the present invention only needs to use 30% of the reagents and a shorter processing time, and the sensitivity and signal strength of the present invention are better than those of the traditional method.

[0472] Example 16 WB4

[0473] Based on the bioreactor provided in Example 6, the reaction is as follows:

[0474] The sample is a His-tagged protein with a molecular weight of 7 kd

[0475] The loading amount is

[0476] 1. 10ng His protein

[0477] 2. 5ng His protein

[0478] 3. 2.5ng His protein

[0479] 4. 1.25ng His protein

[0480] 5. 0.6ng His protein

[0481] 6. 0.3ng His protein

[0482] The blocking solution is 5% skim milk powder in PBS

[0483] PBST is PBS with 1 / 1000 Tween 20

[0484] The primary antibody was mouse anti-His at a concentration of 1 mg / ml, using blocking solution at a dilution of 1:3000

[0485] The secondary antibody was goat anti-mouse HRP at a concentration of 1 mg / ml, using blocking solution at a dilution of 1:10000

[0486] Steps of the present invention:

[0487] 1. The temperature is 20-30°C, the membrane area is 7.5*8CM, and the flow rate is set to 20ml / min;

[0488] 2. Add the primary antibody and secondary antibody stock solutions into the corresponding storage pools (as shown in FIG. 15(C), 23-1 is the secondary antibody stock solution pool, 23-2 is the primary antibody stock solution pool, and 143 is the liquid flow direction);

[0489] 3. Pump in 5 ml of blocking solution. When the blocking solution passes through the reservoir, it will carry out the diluted antibody and enter the reactor. Circulate for 1 hour and then pump out.

[0490] 4. Pump in 5 ml of PBST, circulate for 5 min, and pump out for 3 times;

[0491] 5. Pump in TMB colorimetric solution and pump out;

[0492] 6. Check through the observation window and record the results.

[0493] Traditional method:

[0494] 1. Place two films back to back in a container;

[0495] 2. Add 10 ml of blocking solution, shake for 1 hour, and discard the blocking solution;

[0496] 3. Add 10 ml of PBST, shake for 5 minutes, and discard the PBST;

[0497] 4. Repeat step 3 twice;

[0498] 5. Add 10 ml of primary antibody diluent, shake for 1 hour, and discard the primary antibody diluent;

[0499] 6. Add 10 ml of PBST, shake for 5 minutes, and discard the PBST;

[0500] 7. Repeat step 3 three times;

[0501] 8. Add 10 ml of secondary antibody diluent, shake for 1 hour, and discard the secondary antibody diluent;

[0502] 9. Repeat step 3 three times;

[0503] 10. Add TMB colorimetric solution to the container and record the results.

[0504] Figure 15(B) is the result of the traditional method, and Figure 15(A) is the result of the present invention; Figure 15(C) shows the flow direction of the primary antibody and secondary antibody mother solutions added to the corresponding liquid storage tanks; Figure 15(D) shows a comparative analysis of the coloring degree of the 7kd protein in the results of the traditional method and the results of the present invention.

[0505] Table 16

[0506]

[0507]

[0508] As can be seen from FIG. 15 (D) and Table 16, the stripe depth of the present invention is stronger than that of the traditional method, and the coloring degree is increased by 59.26% to 111.13%, indicating that the present invention has a better effect.

[0509] In summary, compared with the traditional method, the present invention only needs to use 20% of the reagents and a shorter processing time, and the signal intensity of the present invention is better than that of the traditional method.

[0510] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A bioreactor, characterized in that: It comprises a power module (1), a control module (2), a liquid processing module (3), a reactor module (4) and a sensor (5); The power supply module (1) comprises a direct current power supply (6) and a switch (7); The control module (2) comprises a system controller (8), an input device (9) and an output device (10); The liquid processing module (3) comprises a valve (11) or a combination of the valves (11), a pump (12) or a combination of the pumps (12) and a sample pool (13); The reactor module (4) comprises a reactor (14); the reactor (14) comprises a reactor frame (15) and a reactor cavity (16) formed by the reactor frame (15); the reactor frame (15) is provided with at least one opening (17), and the opening (17) is connected to the pump (12) or a combination of the pumps (12); the reactor frame (15) comprises a reactor front plate (20) and a reactor rear plate (21); The power module (1) is respectively connected to the control module (2), the liquid processing module (3), and the reactor module (4) via cables (25); The control module (2) is connected to the liquid processing module (3) and the reactor module (4) respectively through cables (25); The liquid treatment module (3) is connected to the reactor module (4) via a pipeline (26); The control module (2) is connected to the reactor module (4) via the sensor (5).

2. The bioreactor according to claim 1, characterized in that: The reactor frame (15) is also provided with a pressing component (18).

3. The bioreactor according to claim 2, characterized in that: The reactor frame (15) is also provided with a sealing component (19).

4. The bioreactor according to claim 3, characterized in that: A limiting component (22) is also provided between the reactor front plate (20) and the reactor rear plate (21).

5. The bioreactor according to claim 4, characterized in that: A mother liquid pool (23) is also provided between the liquid treatment module (3) and the reactor module (4).

6. The bioreactor according to claim 5, characterized in that: At least one flow dividing block (24) is also arranged in the reactor cavity (16).

7. The bioreactor according to claim 6, characterized in that: The number of the reactor (14) is one or more.

8. The bioreactor according to claim 7, characterized in that: When there are multiple reactors (14), they are connected in series or in parallel.

9. The bioreactor according to claim 8, characterized in that: The reactor (14) is a disposable reactor or a reactor that can be used multiple times.

10. The bioreactor according to claim 9, characterized in that: The inner wall of the reactor (14) is subjected to a hydrophobic treatment or a hydrophilic treatment.

11. The bioreactor according to claim 10, characterized in that: The hydrophobic treatment is one or more of silanization and alkylation.

12. The bioreactor according to claim 10, characterized in that: The hydrophilization treatment is one or more of hydroxylation, carboxylation and amination.

13. The bioreactor according to claim 12, characterized in that: The material of the reactor (14) is a metal material or a polymer material. The metal material is selected from stainless steel and aluminum alloy; the polymer material is selected from polypropylene and acrylonitrile-styrene-butadiene copolymer.

14. The bioreactor according to claim 13, characterized in that: The sealing component (19) is made of an elastic material or a material that repels the reaction solution in the bioreactor.

15. The bioreactor according to claim 14, characterized in that: The reactor module (4) further comprises an identification module.

16. The bioreactor according to claim 15, characterized in that: The identification module is RFID.

17. The bioreactor according to claim 16, characterized in that: The liquid processing module (3) comprises one or more sample placement containers, and the sample placement container is a sample pool (13).

18. The bioreactor according to claim 17, characterized in that: The liquid treatment module (3) comprises at least one pump (12), which is unidirectional or bidirectional.

19. The bioreactor according to claim 18, characterized in that: The pump (12) is one or more of a peristaltic pump, a diaphragm pump, a gear pump, and a plunger pump.

20. The bioreactor according to claim 19, characterized in that: The valve (11) is one or more of a diaphragm valve, a pinch valve, and a column valve.

21. The bioreactor according to claim 20, characterized in that: The reactor (14) is connected to the liquid processing module (3) via a pipeline (26) or a one-way valve or a two-way quick interface.

22. The bioreactor according to claim 21, characterized in that: The bioreactor utilizes a pump (12) to circulate the reaction liquid in the reactor (14).

23. The bioreactor according to claim 22, characterized in that: The bioreactor comprises a liquid sensing module.

24. Use of the bioreactor according to any one of claims 1 to 23 in biological sample detection.

25. A method for detecting a biological sample, characterized in that: Based on the bioreactor as described in any one of claims 1 to 23, a carrier carrying the substance to be reacted is placed in the reactor cavity (16), a reaction liquid is placed in the sample pool (13) of the liquid processing module (3), and a control module (2) is started by a power module (1) to control a pump (12) of the liquid processing module (3) or a combination of the pumps (12) to transport the reaction liquid in the sample pool (13) to the reactor cavity (16) to mix with the carrier carrying the substance to be reacted, react, and detect.

26. The detection method according to claim 25, characterized in that The object to be reacted is a biomacromolecule carrier, and the biomacromolecule carrier is one or more of a film, a polyacrylamide gel, and an agarose gel.

27. The detection method according to claim 26, characterized in that When the reactant is a thin film, the reactor (14) forms a micro cavity.

28. The detection method according to claim 27, characterized in that The biomacromolecule of the biomacromolecule carrier is one or more of nucleic acid, protein and polypeptide.

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