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KL-6 determination reagent and preparation method thereof

A sugar chain antigen and reagent technology, which is applied in the field of salivary liquefied sugar chain antigen determination reagents and its preparation, can solve the problems of lack of detection conditions, inability to accurately quantify, and inability to reflect the severity of the disease, disease discovery and treatment, etc. High accuracy, high accuracy and easy operation

Inactive Publication Date: 2017-06-13
苏州普瑞斯生物科技有限公司
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  • Abstract
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AI Technical Summary

Problems solved by technology

Chromatographic qualitative and enzyme immunoquantitative detection methods are currently available. Chromatographic qualitative can only be used to determine the presence or absence of samples after the concentration of the sample exceeds the detection limit. It cannot be accurately quantified, and cannot reflect the severity of the disease and the early detection and treatment of the disease. , and there are large false negatives or false positives. The enzyme immunoassay method is mainly used by scientific researchers in scientific research. Due to the preparation of detection reagents, there is no unified formula and process, and there is no in-depth study of detection conditions, resulting in different batches. The test results of different reagents are quite different

Method used

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preparation example Construction

[0040] The preparation method of the pretreatment reagent comprises the following steps: mixing a buffer, a signal enhancer, a preservative, an inorganic salt and water to obtain a solution with a pH value of 7.2-7.6;

[0041] The preparation method of the antibody latex reagent comprises the following steps in sequence:

[0042] S1. Dialyzing the anti-saliva glycosylated sugar chain antigen antibody in a sodium bicarbonate buffer solution with a pH of 8.0-9.0 at 2-8 degrees Celsius for at least two times to remove impurities that interfere with the labeling reaction;

[0043] S2. Dilute the anti-saliva liquefied sugar chain antigen antibody solution after dialysis to 1-2 mg / ml with a phosphate buffer solution of pH 9.0-9.5, add an appropriate amount of fluorescein, and continue to mix slightly for 1 hour at 25 degrees Celsius, Carrying out a labeling reaction to prepare a fluoresceinated anti-salivary sugar chain antigen antibody solution;

[0044] S3. Dialyzing the solution...

Embodiment 1

[0058] 1. Preparation of Antibody Latex Reagent

[0059] Dialyze 10 mg of anti-saliva liquefied sugar chain antigen antibody in 0.05 mol / L of pH 9.0, 1 L of sodium bicarbonate buffer solution at 2-8 degrees Celsius for 3 times, each time for 1 hour, to remove preservatives and other interference labeling reactions of impurities.

[0060] Dilute the dialyzed antibody solution to 1 mg / ml with 0.05 mol / L phosphate buffer at pH 9.0, dissolve 1 mg of activated fluorescein FITC in 1 ml of ultrapure water, add it to the antibody solution, and mix gently at 25 degrees Celsius 1 hour for the labeling reaction.

[0061] The fluoresceinized anti-salivary sugar chain antigen antibody solution is dialyzed three times in pH 7.5, 0.05mol / L, 5L phosphate buffer solution at 2-8 degrees Celsius to remove unreacted fluorescein.

[0062] Dilute the dialyzed fluoresceinized antibody to 0.2 mg / ml with pH 7.5, 0.01 mol / L phosphate buffer solution and mix it with an equal volume of 0.1% latex micro...

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Abstract

The invention discloses a KL-6 determination reagent. The KL-6 determination reagent comprises a pretreatment reagent, an antibody latex reagent and an adjusting reagent, wherein the pretreatment reagent 7.2-7.6 in pH value comprises a buffer agent, a signal enhancing agent and a preservative, the antibody latex reagent comprises a buffer agent, latex microspheres and a stabilizing agent, the latex microspheres 100-400 nm in diameter combine anti-human KL-6 antibodies, and the adjusting agent comprises quantitative KL-6. The invention further discloses a preparation method of the KL-6 determination reagent. The KL-6 determination reagent and the preparation method thereof have the advantages that through combination of the anti-human KL-6 antibodies and the latex microspheres, the content of KL-6 in human serum can be determined through a latex-enhanced turbidimetric immunoassay; the KL-6 determination reagent is simple and convenient to operate, high in accuracy degree and repeatability and suitable for high-throughput test, can be used on a fully-automatic biochemical analyzer and well conforms to results determined by an enzyme-linked immunosorbent assay method.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a sialolyzed sugar chain antigen assay reagent capable of measuring the content of sialolyzed sugar chain antigens in human serum by a latex-enhanced immune turbidimetric method and a preparation method thereof. Background technique [0002] KL-6 (salivating sugar chain antigen) belongs to the claster9MUCI class, a glycoprotein with a relative molecular weight close to 200,000. It is mainly secreted by proliferating, regenerating or damaged alveolar type II epithelial cells. In normal lung tissue, KL-6 is expressed on type II alveolar cells, respiratory bronchiole epithelial cells and bronchial gland serous cells. [0003] When the body's alveolar epithelial cells suffer a certain degree of damage, on the one hand, type II alveolar epithelial cells proliferate and activate, and the secretion of KL-6 increases rapidly; on the other hand, the permeability of the lung interstitial basem...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/533
Inventor 吴朝晖
Owner 苏州普瑞斯生物科技有限公司
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