Abnormal sugar chain glycoprotein detection reagent and method for preparing the same

Abstract

The application provides an abnormal sugar chain glycoprotein detection reagent and a preparation method thereof, wherein Tris buffer is used as a solvent, and the content of each component in the detection reagent is as follows: 0.5-2 g / L of disperse blue, 0.3-1.35 g / L of mixed agglutinin, 5-20 mL / L of Prionex 10% stabilizer, and 0.1-0.3 g / L of benzalkonium chloride, and the detection reagent is adjusted to pH 8.0-10.0. The detection reagent has a service life of 3 years, and can simultaneously consider broad spectrum and accuracy.

Description

Technical Field

[0001] This invention relates to the field of protein detection reagent technology, and in particular to a detection reagent for abnormal glycan glycoproteins and its preparation method. Background Technology

[0002] Tumor abnormal protein (TAP): refers to "abnormal glycosylation" in tumor tissue. Abnormal sugars on the surface of tumor cells occur when tumors develop. Due to the inactivation of glycosylation modifying enzymes (glycosyltransferases, glycosylases) or the activation of certain enzymes that are active in the embryonic stage but tend to be quiescent in the mature stage, the glycosylation structure on the surface of cells is changed, which leads to the so-called "abnormal glycosylation" of tumor tissue.

[0003] Lectins are sugar-binding proteins that play a crucial role in cell recognition and adhesion responses, primarily promoting cell-to-cell adhesion. Lectins possess more than one sugar-binding site, thus enabling them to participate in cell recognition and adhesion, connecting different cells.

[0004] In clinical applications, the detection of abnormal glycan glycoproteins can be used to assist in the diagnosis of tumors, assess the effectiveness of tumor treatment, and predict tumor prognosis; sometimes, dynamic monitoring of the quantification of abnormal glycan glycoproteins can also be used to determine tumor recurrence and metastasis.

[0005] Currently, there are techniques that utilize the affinity of abnormal glycan glycoproteins for lectins in tumor diagnosis. However, the following problems exist:

[0006] I. The reagents for detecting abnormal glycan glycoproteins have poor stability and short shelf life, resulting in high storage costs;

[0007] Second, the detection reagents for abnormal glycan glycoproteins have poor repeatability and cannot achieve broad-spectrum and accurate detection. Summary of the Invention

[0008] This invention proposes a detection reagent for abnormal glycan glycoproteins and its preparation method. The detection reagent has a shelf life of up to 3 years and can simultaneously achieve both broad spectrum and accuracy.

[0009] The technical solution of the present invention is implemented as follows: an abnormal glycan glycoprotein detection reagent, wherein the detection reagent uses Tris buffer as solvent, and the contents of each component in the detection reagent are as follows: Disperse Blue 0.5-2 g / L, mixed lectin 0.3-1.35 g / L, Prionex 10% stabilizer 5-20 mL / L, benzalkonium chloride 0.1-0.3 g / L, and the detection reagent is adjusted to pH 8.0-10.0.

[0010] The detection reagent incorporates benzalkonium chloride, a cationic surfactant with dispersing and penetrating properties, which makes the abnormal glycan glycoprotein detection reagent more uniform and enhances the staining effect. Furthermore, benzalkonium chloride is a non-oxidizing bactericide with broad-spectrum and highly effective bactericidal capabilities, inhibiting the growth of various microorganisms such as molds.

[0011] The assay reagent contains 10% Prionex stabilizer. Prionex is an aqueous solution of high-purity porcine dermal collagen polypeptide fragments at a volume percentage of 10%, exhibiting excellent protein stabilizing properties. Compared to bovine serum albumin, it does not pose a potential risk of viral contamination. The addition of 10% Prionex stabilizer ensures a shorter staining time while resulting in a clearer, more defined background, easier slide reading, and better staining outcomes.

[0012] Furthermore, the contents of each component in the test reagent are as follows: Disperse Blue 1g / L, Mixed lectin 0.6g / L, Prionex 10% stabilizer 10mL / L, and benzalkonium chloride 0.2g / L.

[0013] Furthermore, the mixed lectin is one or more of the following: concanavalin A lectin, lentil lectin, wheat germ lectin, soybean lectin, L-type red kidney bean lectin, and citrus fruit pollen lectin.

[0014] Furthermore, in the mixed lectins, the mass ratio of concanavalin A lectin, lentil lectin, wheat germ lectin, and soybean lectin is 3:2:4:3.

[0015] Furthermore, in the mixed lectins, the mass ratio of concanavalin A lectin, lentil lectin, wheat germ lectin, soybean lectin, and L-type red kidney bean lectin is 3:2:4:3:3.

[0016] Furthermore, in the mixed lectins, the mass ratio of concanavalin A lectin, lentil lectin, wheat germ lectin, soybean lectin, L-type red kidney bean lectin, and citrus fruit pollen lectin is 3:2:4:3:3:3.

[0017] Furthermore, the disperse blue is one or both of disperse blue CR-E and disperse blue 14. Using a combination of disperse blue CR-E and disperse blue 14 can enhance the coloring effect and improve the dyeing result.

[0018] Furthermore, the dispersed blue includes dispersed blue CR-E at 0.25-1 g / L and dispersed blue 14 at 0.25-1 g / L.

[0019] Furthermore, the dispersed blue comprises dispersed blue CR-E at 0.5 g / L and dispersed blue 14 at 0.5 g / L.

[0020] Furthermore, the concentration of the Tris buffer is 0.01M to 0.05M, and the pH value is 8.0 to 10.0.

[0021] A method for preparing an abnormal glycan glycoprotein detection reagent includes the following steps:

[0022] a. Weigh out Disperse Blue and Benzalkonium Chloride, add Disperse Blue and Benzalkonium Chloride to purified water, stir and mix well to obtain Disperse Blue solution;

[0023] b. Weigh out Tris, dissolve Tris completely in purified water, and adjust the pH to 8.0-10.0 to obtain 0.01M-0.05M Tris buffer;

[0024] c. Weigh the mixed lectin, add the mixed lectin to Tris buffer, stir and mix well to obtain the mixed lectin solution;

[0025] d. Measure out the disperse blue solution and the mixed lectin solution, and mix the disperse blue solution and the mixed lectin solution at a volume ratio of 1:1 to obtain the abnormal glycan glycoprotein staining solution;

[0026] e. Measure the abnormal glycan glycoprotein staining solution and Prionex 10% stabilizer, mix the abnormal glycan glycoprotein staining solution and Prionex 10% stabilizer evenly, filter, and adjust the pH value to 8.0-10.0 to obtain the abnormal glycan glycoprotein detection reagent.

[0027] A method for using an abnormal glycan glycoprotein detection reagent includes the following steps:

[0028] 1) Take a drop of blood sample and smear it evenly on a glass slide, then let it dry;

[0029] 2) Then add the test reagent, lay flat and let it stand to dry;

[0030] 3) Observe the staining results under a microscope.

[0031] The reaction principle of the test reagent: Under the action of mixed lectins, abnormal glycan glycoproteins in the blood are agglomerated into particles. Disperse blue participates in the agglomeration reaction as a staining agent, which facilitates the observation and identification of agglomerated particles.

[0032] The beneficial effects of this invention are:

[0033] The test reagent of this invention exhibits minimal pH fluctuation over three years, with both positive and negative concordance rates not less than 9 / 10. It has a shelf life of three years and demonstrates excellent stability, repeatability, and accuracy. This test reagent requires only one blood sample for testing, saving time and cost, whereas existing test reagents often require testing three blood samples. The test reagent of this invention utilizes dispersible blue and mixed lectins, further optimized with Prionex 10% stabilizer and benzalkonium chloride, achieving a balance between broad spectrum and accuracy. Detailed Implementation

[0034] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0035] Example 1

[0036] A reagent for detecting abnormal glycan proteins, wherein the reagent uses Tris buffer as a solvent, and the contents of each component in the reagent are as follows: Disperse Blue CR-E 0.25 g / L, Disperse Blue 14 0.25 g / L, Concanavalin A lectin 0.075 g / L, Lentil lectin 0.05 g / L, Wheat germ lectin 0.1 g / L, Soybean lectin 0.075 g / L, Prionex 10% stabilizer 5 mL / L, and Benzalkonium chloride 0.1 g / L.

[0037] The preparation method of the abnormal glycan glycoprotein detection reagent in Example 1 is as follows:

[0038] a. Weigh out Disperse Blue CR-E, Disperse Blue 14 and benzalkonium chloride in the selected proportions, add Disperse Blue CR-E, Disperse Blue 14 and benzalkonium chloride to purified water, stir and mix well to obtain Disperse Blue solution;

[0039] b. Accurately weigh Tris according to the selected ratio, completely dissolve the Tris in purified water, add HCl to adjust the pH to 8.0, and obtain 0.01M Tris buffer;

[0040] c. Accurately weigh concanavalin A lectin, lentil lectin, wheat germ lectin and soybean lectin according to the selected ratio, add the concanavalin A lectin, lentil lectin, wheat germ lectin and soybean lectin to the Tris buffer obtained in step b, stir and mix well to obtain a mixed lectin solution.

[0041] d. Accurately measure the disperse blue solution obtained in step a and the mixed lectin solution obtained in step b according to the selected ratio, and mix the disperse blue solution and the mixed lectin solution evenly at a volume ratio of 1:1 to obtain the abnormal glycan glycoprotein staining solution;

[0042] e. Accurately measure the abnormal glycan glycoprotein staining solution and Prionex 10% stabilizer obtained in step d according to the selected ratio, mix the abnormal glycan glycoprotein staining solution and Prionex 10% stabilizer evenly, filter, and adjust the pH value to 8.0 to obtain the abnormal glycan glycoprotein detection reagent.

[0043] Example 2

[0044] A reagent for detecting abnormal glycan proteins, wherein the reagent uses Tris buffer as a solvent, and the contents of each component in the reagent are as follows: Disperse Blue CR-E 0.5 g / L, Disperse Blue 14 0.5 g / L, Concanavalin A lectin 0.15 g / L, Lentil lectin 0.1 g / L, Wheat germ lectin 0.2 g / L, Soybean lectin 0.15 g / L, Prionex 10% stabilizer 10 mL / L, Benzalkonium chloride 0.2 g / L.

[0045] The preparation method of the abnormal glycan glycoprotein detection reagent in Example 2 is as follows:

[0046] a. Weigh out Disperse Blue CR-E, Disperse Blue 14 and benzalkonium chloride in the selected proportions, add Disperse Blue CR-E, Disperse Blue 14 and benzalkonium chloride to purified water, stir and mix well to obtain Disperse Blue solution;

[0047] b. Accurately weigh Tris according to the selected ratio, completely dissolve the Tris in purified water, add HCl to adjust the pH to 9.0, and obtain 0.02M Tris buffer;

[0048] c. Accurately weigh concanavalin A lectin, lentil lectin, wheat germ lectin and soybean lectin according to the selected ratio, add the concanavalin A lectin, lentil lectin, wheat germ lectin and soybean lectin to the Tris buffer obtained in step b, stir and mix well to obtain a mixed lectin solution.

[0049] d. Accurately measure the disperse blue solution obtained in step a and the mixed lectin solution obtained in step b according to the selected ratio, and mix the disperse blue solution and the mixed lectin solution evenly at a volume ratio of 1:1 to obtain the abnormal glycan glycoprotein staining solution;

[0050] e. Accurately measure the abnormal glycan glycoprotein staining solution and Prionex 10% stabilizer obtained in step d according to the selected ratio, mix the abnormal glycan glycoprotein staining solution and Prionex 10% stabilizer evenly, filter, and adjust the pH value to 9.0 to obtain the abnormal glycan glycoprotein detection reagent.

[0051] Example 3

[0052] A reagent for detecting abnormal glycan proteins, wherein the reagent uses Tris buffer as a solvent, and the contents of each component in the reagent are as follows: Disperse Blue CR-E 1g / L, Disperse Blue 14 1g / L, Concanavalin A lectin 0.225g / L, Lentil lectin 0.15g / L, Wheat germ lectin 0.3g / L, Soybean lectin 0.225g / L, Prionex 10% stabilizer 20mL / L, Benzalkonium chloride 0.3g / L.

[0053] The preparation method of the abnormal glycan glycoprotein detection reagent in Example 3 is as follows:

[0054] a. Weigh out Disperse Blue CR-E, Disperse Blue 14 and benzalkonium chloride in the selected proportions, add Disperse Blue CR-E, Disperse Blue 14 and benzalkonium chloride to purified water, stir and mix well to obtain Disperse Blue solution;

[0055] b. Accurately weigh Tris according to the selected ratio, completely dissolve the Tris in purified water, add HCl to adjust the pH to 10.0, and obtain 0.05M Tris buffer;

[0056] c. Accurately weigh concanavalin A lectin, lentil lectin, wheat germ lectin and soybean lectin according to the selected ratio, add the concanavalin A lectin, lentil lectin, wheat germ lectin and soybean lectin to the Tris buffer obtained in step b, stir and mix well to obtain a mixed lectin solution.

[0057] d. Accurately measure the disperse blue solution obtained in step a and the mixed lectin solution obtained in step b according to the selected ratio, and mix the disperse blue solution and the mixed lectin solution evenly at a volume ratio of 1:1 to obtain the abnormal glycan glycoprotein staining solution;

[0058] e. Accurately measure the abnormal glycan glycoprotein staining solution and Prionex 10% stabilizer obtained in step d according to the selected ratio, mix the abnormal glycan glycoprotein staining solution and Prionex 10% stabilizer evenly, filter, and adjust the pH value to 10.0 to obtain the abnormal glycan glycoprotein detection reagent.

[0059] Example 4

[0060] A reagent for detecting abnormal glycan proteins, wherein the reagent uses Tris buffer as a solvent, and the contents of each component in the reagent are as follows: Disperse Blue CR-E 1g / L, Disperse Blue 14 1g / L, Concanavalin A lectin 0.225g / L, Lentil lectin 0.15g / L, Wheat germ lectin 0.3g / L, Soybean lectin 0.225g / L, L-type red kidney bean lectin 0.225g / L, Prionex 10% stabilizer 20mL / L, and Benzalkonium chloride 0.3g / L.

[0061] The preparation method of the abnormal glycan glycoprotein detection reagent in Example 4 is as follows:

[0062] a. Weigh out Disperse Blue CR-E, Disperse Blue 14 and benzalkonium chloride in the selected proportions, add Disperse Blue CR-E, Disperse Blue 14 and benzalkonium chloride to purified water, stir and mix well to obtain Disperse Blue solution;

[0063] b. Accurately weigh Tris according to the selected ratio, completely dissolve the Tris in purified water, add HCl to adjust the pH to 10.0, and obtain 0.05M Tris buffer;

[0064] c. Accurately weigh concanavalin A lectin, lentil lectin, wheat germ lectin, soybean lectin, and L-type red kidney bean lectin according to the selected proportions, add the concanavalin A lectin, lentil lectin, wheat germ lectin, soybean lectin, and L-type red kidney bean lectin to the Tris buffer solution obtained in step b, stir and mix well to obtain a mixed lectin solution;

[0065] d. Accurately measure the disperse blue solution obtained in step a and the mixed lectin solution obtained in step b according to the selected ratio, and mix the disperse blue solution and the mixed lectin solution evenly at a volume ratio of 1:1 to obtain the abnormal glycan glycoprotein staining solution;

[0066] e. Accurately measure the abnormal glycan glycoprotein staining solution and Prionex 10% stabilizer obtained in step d according to the selected ratio, mix the abnormal glycan glycoprotein staining solution and Prionex 10% stabilizer evenly, filter, and adjust the pH value to 10.0 to obtain the abnormal glycan glycoprotein detection reagent.

[0067] Example 5

[0068] A reagent for detecting abnormal glycan proteins, wherein the reagent uses Tris buffer as a solvent, and the contents of each component in the reagent are as follows: Disperse Blue CR-E 1g / L, Disperse Blue 14 1g / L, Concanavalin A lectin 0.225g / L, Lentil lectin 0.15g / L, Wheat germ lectin 0.3g / L, Soybean lectin 0.225g / L, L-type red kidney bean lectin 0.225g / L, Citrus fruit pollen lectin 0.225g / L, Prionex 10% stabilizer 20mL / L, and benzalkonium chloride 0.3g / L.

[0069] The preparation method of the abnormal glycan glycoprotein detection reagent in Example 5 is as follows:

[0070] a. Weigh out Disperse Blue CR-E, Disperse Blue 14 and benzalkonium chloride in the selected proportions, add Disperse Blue CR-E, Disperse Blue 14 and benzalkonium chloride to purified water, stir and mix well to obtain Disperse Blue solution;

[0071] b. Accurately weigh Tris according to the selected ratio, completely dissolve the Tris in purified water, add HCl to adjust the pH to 10.0, and obtain 0.05M Tris buffer;

[0072] c. Accurately weigh concanavalin A lectin, lentil lectin, wheat germ lectin, soybean lectin, L-type red kidney bean lectin, and citrus fruit pollen lectin according to the selected proportions. Add the concanavalin A lectin, lentil lectin, wheat germ lectin, soybean lectin, L-type red kidney bean lectin, and citrus fruit pollen lectin to the Tris buffer solution obtained in step b, stir and mix well to obtain a mixed lectin solution.

[0073] d. Accurately measure the disperse blue solution obtained in step a and the mixed lectin solution obtained in step b according to the selected ratio, and mix the disperse blue solution and the mixed lectin solution evenly at a volume ratio of 1:1 to obtain the abnormal glycan glycoprotein staining solution;

[0074] e. Accurately measure the abnormal glycan glycoprotein staining solution and Prionex 10% stabilizer obtained in step d according to the selected ratio, mix the abnormal glycan glycoprotein staining solution and Prionex 10% stabilizer evenly, filter, and adjust the pH value to 10.0 to obtain the abnormal glycan glycoprotein detection reagent.

[0075] Comparative Example 1

[0076] The composition of the abnormal glycan glycoprotein detection reagent is basically the same as in Example 2, except that Prionex 10% stabilizer was not added.

[0077] Comparative Example 2

[0078] The composition of the abnormal glycan glycoprotein detection reagent is basically the same as in Example 2, except that benzalkonium chloride is not added.

[0079] The performance test results of the abnormal glycan glycoprotein detection reagents of Examples 1-5 and Comparative Examples 1-2 are as follows:

[0080] pH comparison test: The pH values ​​of the abnormal glycan glycoprotein detection reagents of Examples 1-5 and Comparative Examples 1-2 were measured using an acid-base meter, and the pH values ​​were recorded. The results are shown in Table 1.

[0081] Table 1 pH value test data

[0082] time Example 1 Example 2 Example 3 Example 4 Example 5 Comparative Example 1 Comparative Example 2 Month 0 8.0 9.0 10.0 10.0 9.9 9.0 9.0 June 8.0 9.2 9.8 9.9 10.0 8.9 8.7 December 8.1 8.9 9.9 9.8 10.0 8.2 8.0 24th month 8.0 9.0 9.9 10.0 9.8 7.8 7.6 36th month 8.1 9.2 10.0 9.9 9.7 7.7 7.1 42nd month 8.2 8.8 9.9 9.8 9.9 7.4 7.2

[0083] As shown in Table 1, compared with Comparative Examples 1-2, the pH values ​​of the reagents prepared by the formulations of Examples 1-5 were all within the range of 8.0-10.0 after 6 months, with smaller fluctuations and better stability.

[0084] Positive concordance rate comparison test: The abnormal glycan glycoprotein detection reagents of Examples 1-5 and Comparative Examples 1-2 were used to detect 10 positive samples, and the positive concordance rate was calculated. The results are shown in Table 2.

[0085] Table 2. Positive Concordance Rate Test Data

[0086] time Example 1 Example 2 Example 3 Example 4 Example 5 Comparative Example 1 Comparative Example 2 Month 0 10 / 10 10 / 10 9 / 10 9 / 10 10 / 10 10 / 10 9 / 10 June 10 / 10 10 / 10 10 / 10 10 / 10 9 / 10 9 / 10 10 / 10 December 10 / 10 10 / 10 10 / 10 9 / 10 10 / 10 9 / 10 9 / 10 24th month 9 / 10 10 / 10 9 / 10 9 / 10 9 / 10 7 / 10 6 / 10 36th month 10 / 10 10 / 10 9 / 10 10 / 10 9 / 10 5 / 10 6 / 10 42nd month 9 / 10 10 / 10 10 / 10 9 / 10 9 / 10 5 / 10 4 / 10

[0087] As shown in Table 2, compared with Comparative Examples 1-2, the positive detection rate of the reagents prepared by the formulations of Examples 1-5 after 6 months was not less than 9 / 10, indicating better stability.

[0088] Negative concordance rate comparison test: The abnormal glycan glycoprotein detection reagents of Examples 1-5 and Comparative Examples 1-2 were used to detect 10 negative samples, and the negative concordance rate was calculated. The results are shown in Table 3.

[0089] Table 3. Negative Concordance Rate Test Data Table

[0090] time Example 1 Example 2 Example 3 Example 4 Example 5 Comparative Example 1 Comparative Example 2 Month 0 9 / 10 10 / 10 9 / 10 10 / 10 9 / 10 10 / 10 10 / 10 June 9 / 10 10 / 10 10 / 10 9 / 10 9 / 10 10 / 10 9 / 10 December 10 / 10 9 / 10 10 / 10 10 / 10 9 / 10 9 / 10 9 / 10 24th month 10 / 10 10 / 10 9 / 10 9 / 10 10 / 10 8 / 10 7 / 10 36th month 9 / 10 10 / 10 9 / 10 10 / 10 10 / 10 6 / 10 6 / 10 42nd month 10 / 10 10 / 10 10 / 10 10 / 10 9 / 10 4 / 10 4 / 10

[0091] As shown in Table 3, compared with Comparative Examples 1-2, the negative compliance rate of the reagents prepared by the formulations of Examples 1-5 after 6 months was not less than 9 / 10, indicating better stability.

[0092] Repeatability comparison test: Positive sample R1 and negative sample R2 were tested using the abnormal glycan glycoprotein detection reagents of Examples 1-5 and Comparative Examples 1-2. Each sample was tested 10 times, and the positive and negative concordance rates were calculated. The results are shown in Table 4.

[0093] Table 4 Repeatability Test Data

[0094]

[0095] As shown in Table 4, compared with Comparative Examples 1-2, the reagents prepared by the formulations of Examples 1-5 after 6 months showed positive R1 and negative R2, indicating better repeatability.

[0096] Lowest detection limit comparison test: The abnormal glycan glycoprotein detection reagents of Examples 1-5 and Comparative Examples 1-2 were used to detect four samples L1, L2, L3, and L4 with the lowest detection limit, and the concordance rate was calculated.

[0097] Table 5. Test data for the minimum detection limit

[0098] time sample Example 1 Example 2 Example 3 Example 4 Example 5 Comparative Example 1 Comparative Example 2 Month 0 L1-L4 4 / 4 4 / 4 4 / 4 4 / 4 3 / 4 4 / 4 4 / 4 June L1-L4 4 / 4 3 / 4 4 / 4 3 / 4 4 / 4 3 / 4 4 / 4 December L1-L4 3 / 4 4 / 4 3 / 4 3 / 4 3 / 4 1 / 4 2 / 4 24th month L1-L4 4 / 4 3 / 4 4 / 4 3 / 4 3 / 4 2 / 4 2 / 4 36th month L1-L4 4 / 4 4 / 4 3 / 4 3 / 4 3 / 4 3 / 4 2 / 4 42nd month L1-L4 3 / 4 4 / 4 4 / 4 3 / 4 3 / 4 2 / 4 1 / 4

[0099] As shown in Table 5, compared with Comparative Examples 1-2, the reagents prepared by the formulations of Examples 1-5 after 6 months all had a detection compliance rate of no less than 3 / 4, indicating better accuracy.

[0100] As can be seen from the above, pH value, positive concordance rate, negative concordance rate, repeatability and limit of detection were tested at 0, 6, 12, 24, 36 and 42 months respectively. The test results showed that after 6 months, all performance indicators of Examples 1-5 were better than those of Comparative Examples 1-2, indicating that the addition of Prionex 10% stabilizer and benzalkonium chloride in this invention optimized the reaction system and greatly improved the stability of the reagents.

[0101] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A reagent for detecting abnormal glycan glycoproteins, characterized in that: The detection reagent uses Tris buffer as a solvent, and the contents of each component in the detection reagent are as follows: Disperse Blue 0.5-2 g / L, Mixed lectin 0.3-1.35 g / L, Prionex 10% stabilizer 5-20 mL / L, benzalkonium chloride 0.1-0.3 g / L. The detection reagent is adjusted to pH 8.0-10.

0.

2. The abnormal glycan glycoprotein detection reagent according to claim 1, characterized in that: The contents of each component in the test reagent are as follows: Disperse Blue 1g / L, Mixed lectin 0.6g / L, Prionex 10% stabilizer 10mL / L, and benzalkonium chloride 0.2g / L.

3. A reagent for detecting abnormal glycan glycoproteins according to claim 1 or 2, characterized in that: The mixed lectins are one or more of the following: concanavalin A lectin, lentil lectin, wheat germ lectin, soybean lectin, L-type red kidney bean lectin, and citrus fruit pollen lectin.

4. The abnormal glycan glycoprotein detection reagent according to claim 3, characterized in that: In the mixed lectins, the mass ratio of concanavalin A lectin, lentil lectin, wheat germ lectin, and soybean lectin is 3:2:4:

3.

5. The abnormal glycan glycoprotein detection reagent according to claim 3, characterized in that: In the mixed lectins, the mass ratio of concanavalin A lectin, lentil lectin, wheat germ lectin, soybean lectin, and L-type red kidney bean lectin is 3:2:4:3:

3.

6. The abnormal glycan glycoprotein detection reagent according to claim 3, characterized in that: In the mixed lectins, the mass ratio of concanavalin A lectin, lentil lectin, wheat germ lectin, soybean lectin, L-type red kidney bean lectin, and citrus fruit pollen lectin is 3:2:4:3:3:

3.

7. The abnormal glycan glycoprotein detection reagent according to claim 1, characterized in that: The dispersed blue is one or both of dispersed blue CR-E and dispersed blue 14.

8. The abnormal glycan glycoprotein detection reagent according to claim 7, characterized in that: The dispersed blue includes dispersed blue CR-E at 0.25-1 g / L and dispersed blue 14 at 0.25-1 g / L.

9. A reagent for detecting abnormal glycan glycoproteins according to claim 1 or 2, characterized in that: The concentration of Tris buffer is 0.01M to 0.05M, and the pH value is 8.0 to 10.

0.

10. A method for preparing an abnormal glycan glycoprotein detection reagent according to any one of claims 1-9, characterized in that, Includes the following steps: a. Weigh out Disperse Blue and Benzalkonium Chloride, add Disperse Blue and Benzalkonium Chloride to purified water, stir and mix well to obtain Disperse Blue solution; b. Weigh Tris, dissolve Tris completely in purified water, and adjust the pH to 8.0-10.0 to obtain 0.01M-0.05M Tris buffer; c. Weigh the mixed lectin, add the mixed lectin to Tris buffer, stir and mix well to obtain the mixed lectin solution; d. Measure out the disperse blue solution and the mixed lectin solution, and mix the disperse blue solution and the mixed lectin solution at a volume ratio of 1:1 to obtain the abnormal glycan glycoprotein staining solution; e. Measure the abnormal glycan glycoprotein staining solution and Prionex 10% stabilizer, mix the abnormal glycan glycoprotein staining solution and Prionex 10% stabilizer evenly, filter, and adjust the pH value to 8.0-10.0 to obtain the abnormal glycan glycoprotein detection reagent.

Images