A colloidal gold storage buffer with goat anti-mouse secondary antibody

Abstract

The present application relates to the technical field of biology, and particularly relates to a colloidal gold goat anti-mouse secondary antibody storage buffer.The present application discloses a colloidal gold goat anti-mouse secondary antibody storage buffer, which is prepared by mixing the following components: bovine serum albumin, glycerol, an acid buffer, a pH regulator, ammonium sulfate, dextran, Proclin 300 and goat anti-mouse secondary antibody.The present application has the advantages of simple buffer formula, easy access to raw materials, easy amplification, and improved reactivity and stability of the goat anti-mouse secondary antibody without affecting the performance of the chromatographic test strip.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, and in particular to a colloidal gold buffer solution for preserving goat anti-mouse secondary antibodies. Background Technology

[0002] Colloidal gold immunochromatography is a rapid in vitro detection technique that combines chromatographic and immunochromatographic techniques. It emerged in the early 1980s and is a solid-phase labeled immunoassay technique developed after the three major labeling techniques (fluorescein, radioisotopes, and enzymes). Due to its advantages such as convenience, speed, and ease of storage, it has gained increasing attention and is widely used in immunoassays in humans, animals, and plants.

[0003] The basic materials for colloidal gold immunochromatography using a membrane as the solid-phase carrier are: a microporous membrane coated with a known antigen or antibody; after the sample is added, the antibody or antigen in the sample binds to the antigen or antibody coated on the membrane through filtration or capillary action. This reacts with the colloidal gold label to form a visible red band or spot. In colloidal gold test strips, excess gold-labeled antibody is designed; after being intercepted at the T line, the excess flows to the C line, where it is captured by the C-line antibody, forming a gold-labeled antibody-C-line antibody complex, thus appearing as a red color. The formation of the control line is used to determine the validity of the experiment; therefore, the quality of the control line directly affects the overall quality of the test strip. Generally, the corresponding secondary antibody is selected according to the species of the gold-labeled antibody. For murine antibodies, goat anti-mouse is often chosen as the coating for the C line. Therefore, goat anti-mouse is in high demand as a universal viable material for the quality control line of colloidal gold antigen detection, and it is necessary to improve the reactivity of the viable material. In addition, when coating the antibody on the test strip, too much protective protein should not be added, otherwise it will cause the line to spread and affect the appearance. Without adding protective protein, the antibody has poor stability and is easily precipitated and lost. Therefore, it is very necessary to find a preservation buffer that balances high activity and good stability without affecting the appearance of the C line. Summary of the Invention

[0004] In view of this, the present invention provides a colloidal gold goat anti-mouse secondary antibody preservation buffer solution, which not only ensures the stability of the goat anti-mouse secondary antibody and the aesthetic appearance of the C-line, but also improves the reactivity of the bioactive material; it not only improves the quality of the test strip, but also reduces the amount of bioactive material used.

[0005] To achieve the above-mentioned objectives, the present invention provides the following technical solution:

[0006] This invention provides a colloidal gold goat anti-mouse secondary antibody preservation buffer, comprising an acidic buffer, bovine serum albumin, glycerol, ammonium sulfate and dextran, as well as acceptable excipients or additives.

[0007] In some specific embodiments of the present invention, the pH value of the above-mentioned colloidal gold goat anti-mouse secondary antibody preservation buffer is 4.8, 4.9, 5.0, 5.1 or 5.2.

[0008] In some specific embodiments of the present invention, in the above-mentioned colloidal gold goat anti-mouse secondary antibody preservation buffer: the pH value of the acidic buffer is 4.8, 4.9, 5.0, 5.1 or 5.2;

[0009] The concentration of the acidic buffer solution is 0.05 mol / L, 0.06 mol / L, 0.07 mol / L, 0.08 mol / L, 0.09 mol / L, or 0.1 mol / L;

[0010] The acidic buffer solution includes sodium acetate buffer or sodium citrate buffer.

[0011] In some specific embodiments of the present invention, the above-mentioned colloidal gold is preserved in a goat anti-mouse secondary antibody buffer solution:

[0012] The ammonium sulfate concentration is 5 g / L, 6 g / L, 7 g / L, 8 g / L, 9 g / L, 10 g / L, 11 g / L, 12 g / L, 13 g / L, 14 g / L, 15 g / L, 16 g / L, 17 g / L, 18 g / L, 19 g / L, or 20 g / L; and / or

[0013] The molecular weight of the dextran is 2KD, 3KD, 4KD, 5KD, 6KD, 7KD, 8KD, or 9KD; and / or

[0014] The concentration of the dextran is 5 g / L, 6 g / L, 7 g / L, 8 g / L, 9 g / L or 10 g / L.

[0015] In some specific embodiments of the present invention, the excipients or additives in the above-mentioned colloidal gold goat anti-mouse secondary antibody preservation buffer solution include pH adjusters and preservatives;

[0016] The pH adjuster includes glacial acetic acid or citric acid;

[0017] The preservative includes Proclin 300.

[0018] In some specific embodiments of the present invention, the concentration of Proclin300 in the colloidal gold goat anti-mouse secondary antibody preservation buffer is 0.9 μl / ml, 1.0 μl / ml, or 1.1 μl / ml.

[0019] In some specific embodiments of the present invention, the above-mentioned colloidal gold is preserved in a goat anti-mouse secondary antibody buffer solution:

[0020] The bovine serum albumin content is 1 g / L, 2 g / L, 3 g / L, 4 g / L, 5 g / L, 6 g / L, 7 g / L, 8 g / L, 9 g / L, 10 g / L, 11 g / L, 12 g / L, 13 g / L, 14 g / L, 15 g / L, 16 g / L, 17 g / L, 18 g / L, 19 g / L, or 20 g / L; and / or

[0021] The glycerol content is 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20% (v / v).

[0022] The present invention also provides a colloidal gold goat anti-mouse secondary antibody solution, comprising goat anti-mouse secondary antibody and the above-mentioned colloidal gold goat anti-mouse secondary antibody preservation buffer solution;

[0023] The content of the goat anti-mouse secondary antibody is 10 mg / ml, 15 mg / ml, 20 mg / ml, 25 mg / ml, 30 mg / ml, 35 mg / ml or 40 mg / ml.

[0024] This invention also provides a method for preparing a colloidal gold-based goat anti-mouse secondary antibody solution, comprising the following steps:

[0025] Step (1): Mix ammonium sulfate, dextran and acidic buffer to obtain base buffer, replace goat anti-mouse secondary antibody into the base buffer to obtain antibody solution;

[0026] Step (2): Mix the base buffer, bovine serum albumin, glycerol, and antibody solution to make the bovine serum albumin concentration 1 g / L, 2 g / L, 3 g / L, 4 g / L, 5 g / L, 6 g / L, 7 g / L, 8 g / L, 9 g / L, 10 g / L, 11 g / L, 12 g / L, 13 g / L, 14 g / L, 15 g / L, 16 g / L, 17 g / L, 18 g / L, 19 g / L, or 20 g / L, and the glycerol content 1... The concentrations of the goat anti-mouse secondary antibody are 10 mg / ml, 15 mg / ml, 20 mg / ml, 25 mg / ml, 30 mg / ml, 35 mg / ml, or 40 mg / ml. The solution is filtered and mixed with a preservative to obtain the colloidal gold goat anti-mouse secondary antibody solution.

[0027] In some specific embodiments of the present invention, the above preparation method is as follows:

[0028] The ammonium sulfate concentration is 5 g / L, 6 g / L, 7 g / L, 8 g / L, 9 g / L, 10 g / L, 11 g / L, 12 g / L, 13 g / L, 14 g / L, 15 g / L, 16 g / L, 17 g / L, 18 g / L, 19 g / L, or 20 g / L; and / or

[0029] The pH value of the acidic buffer solution is 4.8, 4.9, 5.0, 5.1, or 5.2; and / or

[0030] The molecular weight of the dextran is 2KD, 3KD, 4KD, 5KD, 6KD, 7KD, 8KD, or 9KD; and / or

[0031] The concentration of the dextran is 5 g / L, 6 g / L, 7 g / L, 8 g / L, 9 g / L, or 10 g / L; and / or

[0032] The concentration of the acidic buffer solution is 0.05 mol / L, 0.06 mol / L, 0.07 mol / L, 0.08 mol / L, 0.09 mol / L, or 0.1 mol / L; and / or

[0033] The acidic buffer solution includes sodium acetate buffer or sodium citrate buffer; and / or

[0034] The preservative includes Proclin 300; and / or

[0035] The concentration of Proclin300 in the colloidal gold goat anti-mouse secondary antibody solution is 0.9 μl / ml, 1.0 μl / ml, or 1.1 μl / ml.

[0036] In some specific embodiments of the present invention, the replacement in the above preparation method includes dialysis replacement or hollow fiber replacement.

[0037] The colloidal gold goat anti-mouse secondary antibody preservation buffer of the present invention has the following effects:

[0038] Goat anti-mouse antibodies are a common active material for colloidal gold control lines, and the demand is very high. Through buffer system research, a buffer that can improve antibody activity and reduce dosage has been identified. In addition, when antibodies are coated on test strips, too much protective protein should not be added, otherwise it will cause the lines to spread. Without the addition of protective protein, the antibody has poor stability and is prone to precipitation. This invention provides a preservation buffer that balances high activity and good stability, and the amount of added protective protein is controlled within an acceptable range for control line interference. Attached Figure Description

[0039] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the accompanying drawings used in the description of the embodiments or the prior art will be briefly introduced below.

[0040] Figure 1 C-line detection is shown. Detailed Implementation

[0041] This invention discloses a colloidal gold-based goat anti-mouse secondary antibody preservation buffer solution. Those skilled in the art can refer to the content of this document and appropriately modify the process parameters to achieve the desired result. It is particularly important to note that all similar substitutions and modifications are obvious to those skilled in the art and are considered to be included in this invention. The methods and applications of this invention have been described through preferred embodiments. Those skilled in the art can clearly modify or appropriately change and combine the methods and applications described herein without departing from the content, spirit, and scope of this invention to realize and apply the technology of this invention.

[0042] To overcome the problems of existing technologies, this invention provides a buffer formulation that improves the reactivity of bioactive materials while ensuring the stability of goat anti-mouse secondary antibodies and the aesthetic appearance of the C-line; it also improves the quality of the test strips and reduces the amount of bioactive materials used. This invention introduces ammonium sulfate, dextran, glycerol, and BSA to enhance protein stability. Although the activity enhancement is only applicable to goat anti-mouse secondary antibodies, the solution to the stability problem can be generalized.

[0043] The colloidal gold goat anti-mouse secondary antibody preservation buffer of the present invention comprises the following components:

[0044] It consists of bovine serum albumin, glycerol, acetate buffer, pH adjuster, ammonium sulfate, dextran, preservative Proclin 300, and goat anti-mouse secondary antibody.

[0045] The bovine serum albumin, glycerol, and protein content in the mixture are in a multiple relationship, expressed as a mass ratio or volume ratio. If the antibody content is set at 10–40 mg / ml, the glycerol content at 1–20%, and the bovine serum albumin content at 1–20 g / L, the mass ratio of antibody, glycerol, and bovine serum albumin is 2:1:1. The acetate is sodium acetate buffer with a concentration of 0.05–0.1 mol / L. The pH adjuster is the acid corresponding to the buffer salt. The ammonium sulfate concentration is 5–20 g / L. The Proclin 300 concentration is 0.9–1.1 μl / ml. The dextran molecular weight is 2–9 KD and the concentration is 5–10 g / L. The pH of the mixture is 5.0 ± 0.2. The protein concentration of the goat anti-mouse secondary antibody is not higher than 40 mg / ml.

[0046] Preferably, the buffer formulation comprises: bovine serum albumin concentration of 5 g / L, glycerol concentration of 5%, sodium acetate buffer concentration of 0.05 mol, glacial acetic acid as pH adjuster, ammonium sulfate concentration of 15 g / L, Proclin 300 concentration of 1 μl / ml, dextran molecular weight of 4 KD and concentration of 10 g / L, pH of the mixture of 5.0, and goat anti-mouse secondary antibody protein concentration of 10 mg / ml.

[0047] The bovine serum albumin was prepared by adjusting the concentration of a 100 g / L stock solution.

[0048] No visible precipitation was observed after the sample buffer was placed at 4°C for 7 days, frozen and thawed 7 times at -20°C, and placed at 37°C for 7 days.

[0049] The preparation method of the colloidal gold goat anti-mouse secondary antibody preservation buffer is as follows:

[0050] 1) Prepare a sodium acetate buffer containing ammonium sulfate and dextran at the pH specified above, and replace the purified antibody into the prepared buffer. This can be done by dialysis or by hollow fiber replacement.

[0051] 2) After the replacement is complete, adjust the protein concentration by adding 10% bovine serum and glycerol to the required concentration;

[0052] 3) Mix the above solution thoroughly, filter through a 0.22μm filter membrane, and store with 0.1% P300.

[0053] The solution is thoroughly mixed by magnetic stirring for 10–20 minutes.

[0054] This invention improves the reactivity and stability of goat anti-mouse secondary antibodies by studying the coating buffer for colloidal gold-coated C-lines, while maintaining the aesthetic appearance of the C-lines. The invention introduces ammonium sulfate, dextran, glycerol, and BSA to enhance protein stability. Although the activity enhancement is only applicable to goat anti-mouse secondary antibodies, the solution to the stability issue can be generalized.

[0055] The method of the present invention will be described in more detail below through specific embodiments to facilitate understanding by those skilled in the art. Unless otherwise specified, the raw materials, reagents, consumables, and instruments involved in the present invention are all commercially available products and can be purchased from the market. The experimental methods used are also conventional methods in the art.

[0056] The goat anti-mouse antibody involved in this invention is a self-made secondary antibody, and the immunogen is mouse IgG, which is obtained by purifying mouse serum SPA.

[0057] The sheep anti-chicken antibody involved in this invention is a self-made secondary antibody. The immunogen is chicken IgY, which is obtained by dissolving chicken IgY in egg yolk water and precipitating it with ammonium sulfate.

[0058] The present invention will be further illustrated below with reference to the embodiments:

[0059] Example 1

[0060] 1) Prepare a pH 5.0 0.05M sodium acetate (containing 5 g / L ammonium sulfate and 5 g / L dextran with a molecular weight of 2 KD) buffer to obtain the base buffer. Dialyze the purified goat anti-mouse secondary antibody 1# into this buffer.

[0061] 2) After dialysis, adjust the protein concentration to 11.1 mg / ml with the above-mentioned base buffer. Add 55 μl each of 100 g / L bovine serum albumin and glycerol to each milliliter of solution to make the final serum albumin concentration 5 g / L and the final glycerol concentration 5%.

[0062] 3) Mix the above solution thoroughly. The protein concentration is 10 mg / ml at this point. Filter through a 0.22 μm filter membrane and store in 1 μl / ml P300 solution.

[0063] Example 2

[0064] Same as Example 1, except that:

[0065] The buffer system was 0.1M sodium acetate buffer, pH 5.2, with ammonium sulfate at a concentration of 20 g / L and dextran (molecular weight 9 KD) at a concentration of 10 g / L. The final protein concentration was 40 mg / ml, the final glycerol concentration was 20%, and the final BSA concentration was 20 g / L.

[0066] Comparative Example 1

[0067] Same as Example 1, except that:

[0068] The buffer system was a 0.05M phosphate buffer with a pH of 7.2. The concentration of ammonium sulfate in the buffer was 15 g / L, and the concentration of dextran (molecular weight 4 KD) in the buffer was 8 g / L. The final protein concentration was 10 mg / ml, the final glycerol concentration was 5%, and the final BSA concentration was 5 g / L.

[0069] Comparative Example 2

[0070] Same as Example 1, except that:

[0071] The buffer system was 0.05M sodium acetate buffer, pH 5, with ammonium sulfate at a concentration of 15 g / L, and dextran at a molecular weight of 4 kDa and a concentration of 8 g / L. The final protein concentration was 10 mg / ml, the final glycerol concentration was 10%, and the final BSA concentration was 5 g / L.

[0072] Comparative Example 3

[0073] Same as Example 1, except that:

[0074] The buffer system was 0.05M sodium acetate buffer, pH 5, with ammonium sulfate at a concentration of 30 g / L, and dextran at a molecular weight of 4 kDa and a concentration of 8 g / L. The final protein concentration was 10 mg / ml, the final glycerol concentration was 5%, and the final BSA concentration was 5 g / L.

[0075] Comparative Example 4

[0076] Same as Example 1, except that:

[0077] The buffer system was 0.05M sodium acetate buffer, pH 5, with ammonium sulfate at a concentration of 15 g / L and dextran (4 kDa) at a concentration of 8 g / L. The final protein concentration was 10 mg / ml, the final glycerol concentration was 10%, and no BSA was added.

[0078] Comparative Example 5

[0079] Same as Example 1, except that:

[0080] The buffer system was 0.05M sodium acetate buffer, pH 5, with ammonium sulfate at a concentration of 15 g / L, and dextran (molecular weight 15 KD) at a concentration of 20 g / L. The final protein concentration was 10 mg / ml, the final glycerol concentration was 5%, and the final BSA concentration was 5 g / L.

[0081] Comparative Example 6

[0082] Same as Example 1, except that:

[0083] The buffer system was 0.05M sodium acetate buffer, pH 5, with ammonium sulfate at a concentration of 15 g / L, and dextran (4 kDa) at a concentration of 8 g / L. The final protein concentration was 50 mg / ml, the final glycerol concentration was 25%, and the final BSA concentration was 25 g / L.

[0084] Comparative Example 7

[0085] Same as Example 1, except that: sheep anti-mouse secondary antibody 1# is replaced with sheep anti-chicken secondary antibody.

[0086] Comparative Example 8

[0087] Same as Comparative Example 1, the difference is that: sheep anti-mouse secondary antibody 1# is replaced with sheep anti-chicken secondary antibody.

[0088] Comparative Example 9

[0089] Same as Example 1, except that: sheep anti-mouse secondary antibody 1# is replaced with sheep anti-mouse secondary antibody 2# (different from sheep anti-mouse secondary antibody 1#, sheep anti-mouse secondary antibody 2# comes from a different numbered animal).

[0090] Comparative Example 10

[0091] Similar to Comparative Example 1, the difference is that sheep anti-mouse secondary antibody 1# is replaced with sheep anti-mouse secondary antibody 2# (different from sheep anti-mouse secondary antibody 1#, sheep anti-mouse secondary antibody 2# comes from different numbered animals).

[0092] Comparative Example 11

[0093] Same as Example 1, except that: ammonium sulfate is replaced with sodium chloride, glycerol is replaced with sorbitol, dextran is replaced with PEG6000, BSA is replaced with HSA, and the concentrations remain the same.

[0094] Comparative Example 12

[0095] Same as Example 1, except that, considering cost, ammonium sulfate, dextran, glycerol, and BSA are added simultaneously in the final product.

[0096] Example of results: Performance evaluation

[0097] The condition parameters of Examples 1-2 and Comparative Examples 1-6 are summarized in Table 1:

[0098] Table 1

[0099]

[0100] 1. Physicochemical property analysis: The antibodies of Examples 1 and 2 and Comparative Examples 1 to 12 were divided into three parts, and placed at 4°C for 7 days, frozen and thawed at -20°C 7 times, and placed at 37°C for 7 days respectively. Turbidimetric analysis (Hach 2100Q portable turbidimeter) was performed. Samples with a value ≤15NTU at 4°C, -20°C, and 37°C (no precipitation can be observed with the naked eye) were defined as qualified samples for stability assessment, as shown in Table 2.

[0101] Table 2: Physicochemical Properties Analysis

[0102]

[0103]

[0104] Except for comparative examples 3-6, 11, and 12, which do not meet the requirements, all others meet the requirements.

[0105] The next step is to test the test strips:

[0106] 2. Potency Assay: A portion of the stable sample stored at 4℃ was selected as a representative for verification. The secondary antibody was diluted to 3 mg / ml, streaked overnight, and air-dried. The gold-labeled mouse anti-COVID-19 antibody was diluted to 2 mg / ml with sample diluent and added to the sample pad. The width and color depth of the C-line were observed and compared with the standard colorimetric card to determine the colorimetric grade (for goat anti-chicken IgY detection, chicken IgY was used). See [link to relevant documentation]. Figure 1 Table 3.

[0107] Table 3: C-line detection results

[0108] type Test results type Test results Example 1 10 Comparative Example 7 10 Example 2 10 Comparative Example 8 7+ Comparative Example 1 7+ Comparative Example 9 9+ Comparative Example 2 9+ (Risk of Spread) Comparative Example 10 10

[0109] The comparison of bands in Example 1, Example 2, and Comparative Example 1 shows that, under the same coating concentration, the detection result of goat anti-mouse secondary antibody preserved in acetic acid buffer at pH 5.0 is about 3 points higher than that preserved in phosphate buffer at pH 7.2.

[0110] Comparing Example 1, Comparative Example 2, and Comparative Example 4, the NTU value of Comparative Example 4 is relatively high, indicating that the addition ratio of protective protein BSA and glycerol is limited, and the additives must be complex. Even if the concentration is increased, it will affect the stability of the goat anti-mouse secondary antibody. The band of Comparative Example 2 is wider than that of Examples 1 and 2, indicating that excessive protective protein has a risk of diffusion.

[0111] Compared with Example 1, Comparative Example 3 showed a higher NTU value, indicating that excessively high ammonium sulfate concentration affects the stability of sheep anti-mouse secondary antibodies, and there is a concentration range to control.

[0112] Compared with Example 1, Comparative Example 5 showed a higher NTU value, indicating that salt concentration, dextran molecular weight and concentration affect the stability of sheep anti-mouse secondary antibody, and higher values ​​are not necessarily better.

[0113] The comparison of NTU values ​​in Examples 1, 2, and 6 shows that it is better to control the antibody concentration below 40 mg / ml, as too high a concentration will affect the stability of the antibody.

[0114] Comparing Examples 1, 1, and 7-10, the turbidity difference was not significant, indicating that this type of additive improves the stability of different proteins. The C-line detection results of Comparative Examples 9 and 10 showed almost no difference, indicating that this buffer system, which improves titer, is only suitable for sheep anti-mouse secondary antibodies.

[0115] In Example 1 and Comparative Example 11, glycerol, sorbitol, BSA, and HSA are similar additives. Sorbitol + BSA is less stable than glycerol + BSA, indicating that glycerol + BSA is the optimal protective protein for sheep anti-mouse secondary antibodies.

[0116] Example 1 and Comparative Example 12 illustrate that the order of additive addition also affects protein stability.

[0117] In summary, the optimal formulation for preserving goat anti-mouse secondary antibody buffer is as follows: bovine serum albumin, glycerol, and protein are in a ratio (if the antibody concentration is set at 10 mg / ml, the glycerol content is 5%, and the bovine serum albumin content is 5 g / L), the acetate is sodium acetate buffer with a concentration of 0.05–0.1 mol / L, the ammonium sulfate concentration is 5–20 g / L, the dextran molecular weight is 2–9 KD with a concentration of 5–10 g / L, the Proclin 300 concentration is 0.9–1.1 μl / ml, the pH of the mixture is 5.0 ± 0.2, and the protein concentration of the goat anti-mouse secondary antibody is not higher than 40 mg / ml. Optimal formulation: antibody content 10mg / ml, glycerol content 5%, bovine serum albumin content 5g / L, sodium acetate buffer concentration 0.05mol / L, ammonium sulfate concentration 15g / L, dextran concentration 4KD, concentration 8g / L, Proclin 300 concentration 1μl / ml, pH of the mixture 5.0.

[0118] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A buffer solution for preserving goat anti-mouse secondary antibody for colloidal gold immunochromatography, characterized in that, It consists of acidic buffer solution, bovine serum albumin, glycerol, ammonium sulfate and dextran, as well as acceptable excipients or adjuvants; The concentration of the acidic buffer solution is 0.05–0.1 mol / L; The content of bovine serum albumin is 1–20 g / L; The content of glycerol is 1% to 20% (v / v); The concentration of ammonium sulfate is 5–20 g / L; The molecular weight of the dextran is 2–9 kDa; The concentration of the dextran is 5–10 g / L; The excipients or additives include preservatives.

2. The colloidal gold immunochromatographic assay buffer solution for preserving goat anti-mouse secondary antibody as described in claim 1, characterized in that, Its pH value is 5.0±0.

2.

3. The colloidal gold immunochromatographic assay buffer solution for preserving goat anti-mouse secondary antibody as described in claim 1 or 2, characterized in that, The pH value of the acidic buffer solution is 5.0 ± 0.2; The acidic buffer solution includes sodium acetate buffer or sodium citrate buffer.

4. The colloidal gold immunochromatographic assay buffer solution for preserving goat anti-mouse secondary antibody as described in claim 1, characterized in that, The excipients or additives also include pH adjusters; The pH adjuster includes glacial acetic acid or citric acid; The preservative includes Proclin 300.

5. The colloidal gold immunochromatographic assay buffer solution for preserving goat anti-mouse secondary antibody as described in claim 4, characterized in that, The concentration of Proclin300 is 0.9–1.1 μl / ml.

6. A goat anti-mouse secondary antibody solution for colloidal gold immunochromatography, characterized in that, Includes goat anti-mouse secondary antibody and the goat anti-mouse secondary antibody preservation buffer for colloidal gold immunochromatography as described in any one of claims 1 to 5; The content of the goat anti-mouse secondary antibody is 10-40 mg / ml.

7. The method for preparing goat anti-mouse secondary antibody solution for colloidal gold immunochromatography as described in claim 6, characterized in that, Includes the following steps: Step (1): Mix ammonium sulfate, dextran and acidic buffer to obtain base buffer, replace goat anti-mouse secondary antibody into the base buffer to obtain antibody solution; Step (2): Take the base buffer, bovine serum albumin, glycerol and the antibody solution and mix them to make the concentration of bovine serum albumin 1-20 g / L, the content of glycerol 1-20% (v / v) and the concentration of goat anti-mouse secondary antibody 10-40 mg / ml. Filter and mix with preservative to obtain the goat anti-mouse secondary antibody solution for colloidal gold immunochromatography.

8. The preparation method according to claim 7, characterized in that, The pH of the acidic buffer solution is 5.0 ± 0.2; the acidic buffer solution includes sodium acetate buffer or sodium citrate buffer; the preservative includes Proclin 300, and the concentration of Proclin 300 in the goat anti-mouse secondary antibody solution for colloidal gold immunochromatography is 0.9–1.1 μl / ml.

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