Confining liquid, preparation method of protein chip and method for improving point-shaped trailing of protein chip

Abstract

The invention relates to a confining liquid, a preparation method of a protein chip and a method for improving point-shaped trailing of the protein chip. The confining liquid comprises the following components: 0.1% v / v-0. 5% v / v of triton X-100, 10 g / L-100 g / L of cane sugar, 0.1 mol / L-0. 2 mol / L of ammonium sulfate, 0.01% v / v-0. 1% v / v of a biological buffer solution, 5% v / v-15% v / v of goat serum and 0.05 g / L-1 g / L of protamine, and the pH value of the confining liquid is 6.5-7.5. After the sample application of the protein chip is completed, the chip is sealed by using the sealing liquid, so that the point shape trailing of the protein chip can be improved.

Description

Technical Field

[0001] The present application relates to the technical field of immunoassay, and in particular to a method for preparing a blocking solution and a protein chip, and a method for improving the spot tailing of a protein chip. Background Art

[0002] Biochip technology is a new type of molecular detection technology that has gradually developed in the early 1990s. It is a micro-biochemical analysis system that integrates discontinuous analysis processes in the field of life sciences on the surface of silicon chips or glass chips based on the principle of specific interactions between molecules through micro-technology to achieve accurate, rapid, and large-volume detection of cells, proteins, genes, and other biological components. Using biochip technology, multiple indicators of the object being tested can be tested at one time. Among them, it is divided into gene chips, protein chips, micro-laboratories, etc.

[0003] Protein chip is an emerging technology that is developing. It provides a universal platform to characterize thousands of proteins in a highly parallel and high-throughput manner. Known proteins are used as probes and fixed on a solid phase carrier using a microarray spotting method. Antibodies or antigens are labeled with markers such as fluorescent substances, enzymes or chemiluminescent substances, react with the probes on the chip, and bind to the detected protein through protein-protein interactions. The hybridization results are then scanned by eye or through a specific device, and finally qualitative or quantitative analysis is performed.

[0004] The immunological method for detecting marker antibodies is an indirect method. The antigen that fixes the capture antibody is a chip matrix with glass as the carrier. The antigen of each marker is diluted with the spotting solution and sprayed to the corresponding position in a matrix in sequence using a piezoelectric spotter. These antigens diluted with the spotting solution can capture specific antibodies in the sample to be tested. The captured antibodies combine with the second antibody labeled with HRP to form a complex. HRP can catalyze the chemiluminescent substrate, and its light signal is collected by a CCD camera. The strength of the light signal can be used to determine the concentration of specific marker antibodies in the sample to be tested.

[0005] Protein chips are widely used in biomedical research, but during the test process, some indicators may have point tailing problems, causing abnormal results. The blocking method in the prior art fails to effectively solve this problem, so a method is needed to improve the quality and stability of the chip's point pattern. Summary of the invention

[0006] Based on this, it is necessary to provide a sealing liquid in one embodiment of the present application to improve the quality and stability of the dot pattern of the chip.

[0007] The technical solutions include the following:

[0008] A blocking solution for improving the tailing phenomenon of a protein chip, wherein the blocking solution comprises components of Triton X-100 0.1% v / v to 0.5% v / v, sucrose 10 g / L to 100 g / L, ammonium sulfate 0.1 mol / L to 0.2 mol / L, biological buffer 0.01% v / v to 0.1% v / v, goat serum 5% v / v to 15% v / v and protamine 0.05 g / L to 1 g / L, and the pH value is 6.5 to 7.5.

[0009] In one embodiment, the components of the blocking solution include Triton X-100 0.1% v / v to 0.2% v / v, sucrose 50 g / L to 100 g / L, ammonium sulfate 0.1 mol / L to 0.15 mol / L, biological buffer 0.05% v / v to 0.1% v / v, goat serum 5% v / v to 10% v / v and protamine 0.25 g / L to 1 g / L.

[0010] In one embodiment, the biological buffer comprises a phosphate buffer.

[0011] In one embodiment, the concentration of phosphate in the phosphate buffer is 0.5 mmol / L~5 mmol / L.

[0012] In one embodiment, the phosphate is disodium hydrogen phosphate.

[0013] The method for preparing the blocking solution comprises: mixing the Triton X-100, sucrose, ammonium sulfate, biological buffer, goat serum and protamine with water to prepare the blocking solution.

[0014] A method for preparing a protein chip, comprising:

[0015] Spot the antigen or antibody on the chip carrier and let it stand;

[0016] The chip carrier after standing is sealed with the sealing solution to prepare a protein chip.

[0017] In one embodiment, the standing time is 0.5h to 1h; and / or the sealing time is 12h to 20h.

[0018] A kit comprises a protein chip prepared by the preparation method.

[0019] In one embodiment, the method further comprises a secondary antibody labeled with an enzyme and an enzymatic chemiluminescent agent.

[0020] The protein chip prepared by the preparation method is used in preparing products for detecting antigens or antibodies.

[0021] A method for improving the spot tailing of a protein chip, comprising:

[0022] The protein chip prepared by the preparation method is subjected to immune reaction with the sample to be tested, and the image is captured by a biochip analyzer.

[0023] In one embodiment, the method is based on chemiluminescence. Optionally, the chemiluminescence method includes enzymatic chemiluminescence.

[0024] In one embodiment, the enzymatic chemiluminescence method includes subjecting the protein chip to an immunological reaction with a sample to be tested, adding a second antibody labeled with an enzyme to react after the reaction, and adding an enzymatic chemiluminescent agent after the reaction.

[0025] Compared with the traditional technology, this application has the following beneficial effects:

[0026] The present application combines a surfactant, Triton X-100, a protective agent, sucrose, an ammonium sulfate solution, a biological buffer, goat serum and protamine to form a blocking solution of a specific formula. After the chip carrier is spotted, the chip carrier after the spotting is blocked with the blocking solution, which can improve the spot tailing problem of the protein chip, thereby improving the accuracy of the immunoassay reagent.

[0027] Based on the above-mentioned blocking solution, a method for improving the dot tailing of a protein chip is provided, thereby improving the dot pattern quality and stability of the chip, thereby improving the accuracy of the immunoassay reagent.

[0028] Furthermore, after the protein chip is spotted, the standing time and / or blocking time are adjusted to further improve the spot tailing of the protein chip. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] In order to more clearly illustrate the technical solutions in the embodiments of the present application and to more completely understand the present application and its beneficial effects, the following is a brief introduction to the drawings required for the description of the embodiments. Obviously, the drawings described below are only some embodiments of the present application, and those skilled in the art can obtain other drawings based on these drawings without creative work.

[0030] Figure 1 The test results of the protein chip blocked with the blocking solution of Example 1;

[0031] Figure 2 The test results of the protein chip blocked with the blocking solution of Example 2;

[0032] Figure 3 The test results of the protein chip blocked with the blocking solution of Example 3;

[0033] Figure 4 The test results of the protein chip blocked with the blocking solution of Example 4 are shown in FIG.

[0034] Figure 5 The test results of the protein chip blocked with the blocking solution of Comparative Example 1 are shown in FIG.

[0035] Figure 6 The test results of the protein chip blocked with the blocking solution of Comparative Example 2 are shown below:

[0036] Figure 7 The test results of the protein chip blocked with the blocking solution of Comparative Example 3 are shown below;

[0037] Figure 8 The test results of the protein chip blocked with the blocking solution of Comparative Example 4 are shown below;

[0038] Fig. 9 The test results of the protein chip blocked with the blocking solution of Comparative Example 5 are shown below;

[0039] Fig.10 The protein chip test results of Example 5;

[0040] Fig.11 The protein chip test results of Example 6;

[0041] Fig.12 The protein chip test results of Example 7;

[0042] Fig.13 The protein chip test results of Example 8;

[0043] Fig.14 The protein chip test results of Example 9;

[0044] Fig.15 This is the protein chip test result of Example 10. DETAILED DESCRIPTION

[0045] In order to make the above-mentioned purposes, features and advantages of the present application more obvious and easy to understand, the specific implementation methods of the present application are described in detail below. In the following description, many specific details are set forth to facilitate a full understanding of the present application. However, the present application can be implemented in many other ways different from those described herein, and those skilled in the art can make similar improvements without violating the connotation of the present application, so the present application is not limited by the specific embodiments disclosed below.

[0046] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which this application belongs. The terms used herein in the specification of this application are only for the purpose of describing specific embodiments and are not intended to limit this application.

[0047] Herein, when it comes to a numerical interval (i.e., a numerical range), unless otherwise specified, the distribution of the optional numerical values ​​within the numerical interval is considered to be continuous and includes the two numerical endpoints (i.e., the minimum and maximum values) of the numerical interval, and each numerical value between the two numerical endpoints. Unless otherwise specified, when a numerical interval only refers to integers within the numerical interval, including the two endpoint integers of the numerical range, and each integer between the two endpoints, is equivalent to directly listing each integer. When multiple numerical ranges are provided to describe features or characteristics, these numerical ranges can be combined. In other words, unless otherwise specified, the numerical ranges disclosed herein should be understood to include any and all sub-ranges included therein. The "numerical value" in the numerical interval can be any quantitative value, such as a number, a percentage, a ratio, etc. "Numerical interval" allows broadly including numerical interval types such as percentage intervals, ratio intervals, and ratio intervals.

[0048] In this article, if there are multiple steps involved in the method flow, unless there is a clear different description in this article, there is no strict order restriction for the execution of these steps, and they can be executed in other orders than described. Moreover, any step can include multiple sub-steps or multiple stages, and these sub-steps or stages are not necessarily executed at the same time, but can be executed at different times, and their execution order is not necessarily sequential, but can be executed in turn, alternating or simultaneously with other steps or parts of sub-steps or stages of other steps.

[0049] The terms "and / or", "or / and", and "and / or" used herein include any one of two or more related listed items, and also include any and all combinations of the related listed items. "Any and all combinations" include any combination of two related listed items, any more related listed items, or all related listed items. For example, "A and / or B" includes three parallel solutions: A, B, and "a combination of A and B".

[0050] In this document, "multiple", "multiple", "multiple times", "multiple", "several", etc., unless otherwise specified, refer to a number greater than or equal to 2. For example, "one or more" means one or greater than or equal to two.

[0051] Herein, "preferred", "preferable", "better", etc. are only used to describe implementation methods or examples with better effects. It should be understood that they do not constitute limitations on the scope of protection of this application.

[0052] One embodiment of the present application provides a blocking solution, the components of which include surfactant Triton X-100 0.1% v / v~0.5% v / v, protective agent sucrose 10g / L~100g / L, ammonium sulfate 0.1mol / L~0.2mol / L, biological buffer 0.01%v / v~0.1%v / v, goat serum 5%v / v~15%v / v and protamine 0.05g / L~1g / L, and the pH value is 6.5~7.5. The chip carrier is blocked by the blocking solution, which can improve the point tailing problem of the protein chip, thereby improving the accuracy of the immunoassay reagent.

[0053] In a specific example, the concentration of Triton X-100 in the blocking solution can be, but is not limited to, 0.1% v / v, 0.2% v / v, 0.3% v / v, 0.4% v / v, 0.5% v / v, or a range between any two of the above values.

[0054] In a specific example, the concentration of sucrose in the blocking solution can be but is not limited to 10 g / L, 15 g / L, 20 g / L, 25 g / L, 30 g / L, 35 g / L, 40 g / L, 45 g / L, 50 g / L, 55 g / L, 60 g / L, 65 g / L, 70 g / L, 75 g / L, 80 g / L, 85 g / L, 90 g / L, 95 g / L, 100 g / L, or a range between any two of the above values.

[0055] In a specific example, the concentration of ammonium sulfate in the blocking solution can be, but is not limited to, 0.1 mol / L, 0.12 mol / L, 0.15 mol / L, 0.18 mol / L, 0.2 mol / L, or a range between any two of the above values.

[0056] In a specific example, the concentration of the biological buffer in the blocking solution can be, but is not limited to, 0.01% v / v, 0.02% v / v, 0.03% v / v, 0.04% v / v, 0.05% v / v, 0.06% v / v, 0.07% v / v, 0.08% v / v, 0.09% v / v, 0.1% v / v, or a range between any two of the above values.

[0057] In a specific example, the concentration of goat serum in the blocking solution can be but is not limited to 5% v / v, 6% v / v, 7% v / v, 8% v / v, 9% v / v, 10% v / v, 11% v / v, 12% v / v, 13% v / v, 14% v / v, 15% v / v, or a range between any two of the above values.

[0058] In a specific example, the concentration of protamine in the blocking solution can be, but is not limited to, 0.05 g / L, 0.1 g / L, 0.15 g / L, 0.2 g / L, 0.25 g / L, 0.3 g / L, 0.35 g / L, 0.4 g / L, 0.45 g / L, 0.5 g / L, 0.55 g / L, 0.6 g / L, 0.65 g / L, 0.7 g / L, 0.75 g / L, 0.8 g / L, 0.85 g / L, 0.9 g / L, 0.95 g / L, 1 g / L, or a range between any two of the above values.

[0059] In a specific example, the components of the blocking solution include a surfactant of 0.1% v / v to 0.2% v / v, a protective agent of 50 g / L to 100 g / L, ammonium sulfate of 0.1 mol / L to 0.15 mol / L, a biological buffer of 0.05% v / v to 0.1% v / v, goat serum of 5% v / v to 10% v / v, and protamine of 0.25 g / L to 1 g / L.

[0060] In a preferred specific example, the components of the blocking solution include 0.1% v / v surfactant, 100 g / L protective agent, 0.1 mol / L ammonium sulfate, 0.1% v / v biological buffer, 10% v / v goat serum and 1 g / L protamine. The blocking solution has a better effect of improving the spot tailing of the protein chip.

[0061] In one specific example, the biological buffer includes a phosphate buffer.

[0062] In a specific example, the concentration of phosphate in the phosphate buffer is 0.5 mmol / L to 5 mmol / L.

[0063] In a specific example, the phosphate buffer is a disodium hydrogen phosphate solution, and the concentration of disodium hydrogen phosphate in the disodium hydrogen phosphate solution is 0.5 mol / L to 5 mol / L. Optionally, the concentration of disodium hydrogen phosphate in the disodium hydrogen phosphate solution is 0.5 mol / L, 1 mol / L, 1.5 mol / L, 2 mol / L, 2.5 mol / L, 3 mol / L, 3.5 mol / L, 4 mol / L, 4.5 mol / L, 5.0 mol / L, or a range consisting of any two of the above values.

[0064] One embodiment of the present application further provides a method for preparing the above-mentioned blocking solution, comprising: mixing Triton X-100, sucrose, ammonium sulfate, biological buffer, goat serum and protamine with water to prepare the blocking solution.

[0065] One embodiment of the present application also provides a method for preparing a protein chip, comprising steps S1-S2:

[0066] Step S1, spot the antigen or antibody on the chip carrier and let it stand.

[0067] In a specific example, the standing time is 0.5h to 2h, and optionally, the standing time is 0.5h, 1h, 1.5h, 2h, or a range consisting of any two of the above values. Preferably, the standing time is 1h, which has a better effect on improving the spot tailing of the protein chip.

[0068] In one specific example, the antigen or antibody is a diluted antigen or antibody.

[0069] In a specific example, the material of the chip substrate is siliconized glass.

[0070] Step S2, using the blocking solution to block the chip carrier after standing to prepare a protein chip.

[0071] In a specific example, the protein chip is dropped vertically into the blocking solution and maintained for a specified blocking time.

[0072] In a specific example, the blocking time is 12h to 20h, and optionally, the standing time is 12h, 13h, 14h, 15h, 16h, 17h, 18h, 19h, 20h, or a range consisting of any two of the above values. Preferably, the blocking time is 16h, which has a better effect on improving the spot tailing of the protein chip.

[0073] One embodiment of the present application further provides a kit, comprising a protein chip prepared by the preparation method.

[0074] In a specific example, a secondary antibody labeled with an enzyme and an enzymatic chemiluminescent agent are also included.

[0075] One embodiment of the present application also provides the use of the protein chip prepared by the above preparation method or the above kit in the preparation of products for detecting antigens or antibodies.

[0076] One embodiment of the present application also provides a method for improving the spot tailing of a protein chip, comprising:

[0077] Based on the chemiluminescence method, the protein chip prepared by the preparation method is subjected to immune reaction with the sample to be tested, and the image is captured by a biochip analyzer.

[0078] In one specific example, the chemiluminescence method includes enzymatic chemiluminescence method.

[0079] In a specific example, the enzymatic chemiluminescence method includes subjecting the protein chip to an immunological reaction with a sample to be tested, adding a second antibody labeled with an enzyme to react after the reaction, and adding an enzymatic chemiluminescent agent after the reaction.

[0080] Optionally, the enzyme comprises horseradish peroxidase and / or alkaline phosphatase.

[0081] The embodiments of the present application will be described in detail below in conjunction with examples. It should be understood that these examples are only used to illustrate the present application and are not intended to limit the scope of the present application. The experimental methods for which specific conditions are not specified in the following examples are preferably referred to the guidance provided in the present application, and can also be based on the experimental manual or normal conditions in this area, can also be based on the conditions recommended by the manufacturer, or refer to experimental methods known in the art.

[0082] In the following specific embodiments, the measured parameters of raw material components may have slight deviations within the range of weighing accuracy unless otherwise specified. For temperature and time parameters, acceptable deviations caused by instrument test accuracy or operation accuracy are allowed.

[0083] The biochip analyzer used in the following examples and comparative examples is a Sanlian Biotech fully automatic biochip analyzer SLXP-001B, and the phosphate buffer is a 5 mol / L disodium hydrogen phosphate solution.

[0084] Example 1

[0085] This embodiment provides a sealing solution formula and a preparation method thereof:

[0086] The blocking solution includes 0.1% surfactant, 10 g / L protective agent, 0.1 mol / L ammonium sulfate, 0.01% biological buffer, 5% goat serum and 0.05 g / L protamine, with a pH value of 6.5. The specific formula is shown in Table 1.

[0087] Preparation method: Use purified water as the matrix, mix with the above-mentioned surfactant, protective agent, ammonium sulfate, biological buffer, goat serum and protamine, and mix thoroughly.

[0088] Example 2

[0089] This embodiment provides a sealing solution formula and a preparation method thereof:

[0090] The blocking solution includes 0.2% surfactant, 50 g / L protective agent, 0.15 mol / L ammonium sulfate, 0.05% biological buffer, 10% goat serum and 0.25 g / L protamine, with a pH value of 7.0. The specific formula is shown in Table 1.

[0091] Preparation method: Use purified water as the matrix, mix with the above-mentioned surfactant, protective agent, ammonium sulfate, biological buffer, goat serum and protamine, and mix thoroughly.

[0092] Example 3

[0093] This embodiment provides a sealing solution formula and a preparation method thereof:

[0094] The blocking solution includes 0.5% surfactant, 100 g / L protective agent, 0.2 mol / L ammonium sulfate, 0.1% biological buffer, 15% goat serum and 0.5 g / L protamine, with a pH value of 7.5. The specific formula is shown in Table 1.

[0095] Preparation method: Use purified water as the matrix, mix with the above-mentioned surfactant, protective agent, ammonium sulfate, biological buffer, goat serum and protamine, and mix thoroughly.

[0096] Example 4

[0097] This embodiment provides a sealing solution formula and a preparation method thereof:

[0098] The blocking solution includes 0.1% surfactant, 100 g / L protective agent, 0.1 mol / L ammonium sulfate, 0.1% biological buffer, 10% goat serum and 1 g / L protamine, with a pH value of 6.5. The specific formula is shown in Table 1.

[0099] Preparation method: Use purified water as the matrix, mix with the above-mentioned surfactant, protective agent, ammonium sulfate, biological buffer, goat serum and protamine, and mix thoroughly.

[0100] Table 1

[0101]

[0102] Comparative Example 1

[0103] This comparative example provides a sealing solution formula and a preparation method thereof, which is different from Example 4 in that Triton X-100 is replaced by CA-630, and other contents are the same.

[0104] Comparative Example 2

[0105] This comparative example provides a sealing solution formula and a preparation method thereof, which is different from Example 4 in that Triton X-100 is replaced by NP-40, and other contents are the same.

[0106] Comparative Example 3

[0107] This comparative example provides a sealing solution formula and a preparation method thereof, which is different from Example 4 in that Triton X-100 is replaced by Brij 35, and other contents are the same.

[0108] Comparative Example 4

[0109] This comparative example provides a sealing solution formula and a preparation method thereof, which is different from Example 4 in that sucrose is replaced by trehalose, and other contents are the same.

[0110] Comparative Example 5

[0111] This comparative example provides a sealing solution formula and a preparation method thereof, which is different from Example 4 in that sucrose is replaced by PEG 6000, and other contents are the same.

[0112] test

[0113] During the experiment, since the Scl-70 index in the autoimmune chip had the most serious tailing among all the indicators, the chip with Scl-70 antigen spots was blocked using the blocking solutions of Examples 1 to 4 and Comparative Examples 1 to 5, and the anti-Scl-70 antibodies in human serum or plasma samples were detected, and the chip image results were observed.

[0114] The specific steps include:

[0115] 1. Select a clinical sample with a high concentration of Scl-70 collected and processed according to the correct medical technology for antinuclear antibody testing.

[0116] 2. Blocking Solution The blocking solution was prepared according to the method of each embodiment and comparative example.

[0117] 3. Spot the Scl-70 antigen separately, and after spotting, let the chip stand for 30 minutes, and use different formulas of blocking solutions to block for 20 hours; after reacting with the treated clinical samples, add the sheep anti-human IgG antibody labeled with HRP to react, and finally add the chemiluminescent substrate to observe the pictures after taking pictures with the biochip analyzer. The results of Examples 1 to 4 are as follows: Figure 1~Figure 4 As shown, the results of comparative examples 1 to 5 are as follows Figure 5~Figure 9 shown.

[0118] Embodiment 5~embodiment 7

[0119] The blocking solution formula with the least tailing problem was screened out, and the blocking solution formula of Example 4 was selected. The Scl-70 antigen was spotted separately. After the spotting was completed, according to Examples 5 to 7 as shown in Table 2, it was left to stand for 30 minutes, 1 hour or 2 hours, respectively, and then blocked with the same blocking solution for 16 hours; after reacting with the treated clinical sample, a sheep anti-human IgG antibody labeled with HRP was added for reaction, and finally a chemiluminescent substrate was added to observe the picture after taking it with a biochip analyzer. The results of Examples 5 to 7 are as follows: Figure 10~Figure 12 As shown, compare the chip image results.

[0120] Table 2

[0121]

[0122] Embodiment 8~embodiment 10

[0123] The standing time after spotting with the least tailing problem was screened out, and the Scl-70 antigen was spotted separately. After spotting, it was stood for the same time, and then blocked with the same blocking solution. According to Examples 8 to 10 in Table 3 below, the blocking was performed for 12 hours, 16 hours or 20 hours respectively; after reacting with the treated clinical sample, a sheep anti-human IgG antibody labeled with HRP was added for reaction, and finally a chemiluminescent substrate was added to observe the picture after taking it with a biochip analyzer. The results of Examples 8 to 10 are as follows: Figure 13~Figure 15 As shown, compare the chip image results.

[0124] Table 3

[0125]

[0126] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0127] The above-mentioned embodiments only express several implementation methods of the present application, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the invention patent. It should be pointed out that, for a person of ordinary skill in the art, several variations and improvements can be made without departing from the concept of the present application, and these all belong to the protection scope of the present application. Therefore, the protection scope of the patent of the present application shall be subject to the attached claims, and the description may be used to interpret the content of the claims.

Claims

1. A blocking solution for improving the tailing phenomenon of a protein chip, characterized in that: The components of the blocking solution include Triton X-100 0.1% v / v to 0.5% v / v, sucrose 10 g / L to 100 g / L, ammonium sulfate 0.1 mol / L to 0.2 mol / L, biological buffer 0.01% v / v to 0.1% v / v, goat serum 5% v / v to 15% v / v and protamine 0.05 g / L to 1 g / L, and the pH value is 6.5 to 7.

5.

2. The sealing liquid according to claim 1, characterized in that The components of the blocking solution include Triton X-1000 0.1% v / v to 0.2% v / v, sucrose 50 g / L to 100 g / L, ammonium sulfate 0.1 mol / L to 0.15 mol / L, biological buffer 0.05% v / v to 0.1% v / v, goat serum 5% v / v to 10% v / v and protamine 0.25 g / L to 1 g / L.

3. The sealing liquid according to claim 1 or 2, characterized in that: Biological buffers include phosphate buffer; Optionally, the concentration of phosphate in the phosphate buffer is 0.5 mmol / L to 5 mmol / L; Optionally, the phosphate is disodium hydrogen phosphate.

4. The method for preparing the blocking solution according to any one of claims 1 to 3, characterized in that: include: The Triton X-100, sucrose, ammonium sulfate, biological buffer, goat serum and protamine were mixed with water to prepare a blocking solution.

5. A method for preparing a protein chip, characterized in that: include: Spot the antigen or antibody on the chip carrier and let it stand; The chip carrier after standing is blocked with the blocking solution according to any one of claims 1 to 3 to prepare a protein chip.

6. The preparation method according to claim 5, characterized in that: The standing time is 0.5h to 1h; and / or the sealing time is 12h to 20h.

7. A kit, characterized in that: A protein chip prepared by the preparation method according to any one of claims 5 to 6; Optionally, a secondary antibody labeled with an enzyme and an enzymatic chemiluminescent agent are also included.

8. Use of the protein chip prepared by the preparation method according to any one of claims 5 to 6 in preparing products for detecting antigens or antibodies.

9. A method for improving the spot tailing of a protein chip, characterized in that: include: The protein chip prepared by the preparation method according to any one of claims 5 to 6 is subjected to immune reaction with a sample to be tested, and an image is captured using a biochip analyzer.

10. The method according to claim 9, characterized in that The method is based on chemiluminescence, and optionally, the chemiluminescence comprises enzymatic chemiluminescence; Optionally, the enzymatic chemiluminescence method includes subjecting the protein chip to an immunological reaction with a sample to be tested, adding a second antibody labeled with an enzyme to react after the reaction, and adding an enzymatic chemiluminescent agent after the reaction.

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