Adiponectin-latex enhanced turbidimetric immunoassay kit and preparation method thereof

An immunoturbidimetric and latex-enhanced technology, applied in the biological field, can solve problems such as pollution, low degree of automation, and long operation time, and achieve the effects of increasing linear range, improving sensitivity, and improving repeatability

Inactive Publication Date: 2019-12-13
苏州普瑞斯生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the ELISA method has been used clinically for nearly two decades, it still has some fatal shortcomings, such as long operation time and low degree of automation
Radioimmunoassay (RIA) has high sensitivity, but is unstable and has poorer repeatability than ELISA, and there is a risk of radioactive contamination

Method used

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  • Adiponectin-latex enhanced turbidimetric immunoassay kit and preparation method thereof
  • Adiponectin-latex enhanced turbidimetric immunoassay kit and preparation method thereof
  • Adiponectin-latex enhanced turbidimetric immunoassay kit and preparation method thereof

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preparation example Construction

[0048] A preparation method of adiponectin-latex enhanced immune turbidity kit, characterized in that: comprising the following steps: A), preparation of reagent R1:

[0049] First add part of purified water to the magnetic stirrer in the liquid mixing tank, adjust the stirrer speed to 200-300rpm to keep the stirring at a constant speed, and put in the first buffer solution, sodium chloride, first stabilizer, surface Active agent, polyethylene glycol 6000 and the first preservative, stir until the materials are completely dissolved, the solution is clear and transparent, and there is no sediment at the bottom of the liquid mixing tank, adjust the pH value between 5.90-8.90 and adjust the volume to the final volume;

[0050] The dissolved solution is filtered through the microporous membrane according to the operating procedures of the microporous filter, and the filtered solution is stored in the finished product tank for identification;

[0051] B), preparation of reagent R2:...

Embodiment 1

[0066] A preparation method of adiponectin-latex enhanced immune turbidity kit, comprising the following steps: A), preparation of reagent R1:

[0067] First add part of purified water to the magnetic stirrer in the liquid mixing tank, adjust the stirrer speed to 200rpm to keep the stirring at a constant speed, and put 6.0g / L tris hydroxymethyl aminomethane, 5.0g / L tris Sodium chloride, 0.5g / L casein, 1ml / L Tween 20, 60g / L polyethylene glycol 6000 and 0.8ml / L sodium azide were stirred until the materials were completely dissolved and the solution was clear and transparent. There is no precipitation at the bottom of the liquid tank, adjust the pH value to 5.90 and adjust the volume to the final volume;

[0068] The dissolved solution is filtered through the microporous membrane according to the operating procedures of the microporous filter, and the filtered solution is stored in the finished product tank for identification;

[0069] B), preparation of reagent R2:

[0070] Pr...

Embodiment 2

[0083]A preparation method of adiponectin-latex enhanced immune turbidity kit, comprising the following steps: A), preparation of reagent R1:

[0084] First add part of purified water to the magnetic stirrer in the liquid mixing tank, adjust the stirrer speed to 250rpm to keep the stirring at a constant speed, and add 9.0g / L tris hydroxymethyl aminomethane and 16g / L chlorine Sodium chloride, 1.2g / L mannitol, 3ml / L Triton X-100, 90g / L polyethylene glycol 6000 and 1.4ml / L Proclin-950, stir until the material is completely dissolved and the solution is clear and transparent , There is no precipitation at the bottom of the liquid mixing tank, adjust the pH value to 7.4 and adjust the volume to the final volume;

[0085] The dissolved solution is filtered through the microporous membrane according to the operating procedures of the microporous filter, and the filtered solution is stored in the finished product tank for identification;

[0086] B), preparation of reagent R2:

[00...

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Abstract

The invention belongs to the technical field of biology and relates to a preparation method of an adiponectin-latex enhanced turbidimetric immunoassay kit. A reagent R2 is composed of a reaction solution, a preservation solution and a latex microsphere antibody conjugate; the reaction solution is composed of a second buffer solution; and the latex microsphere antibody conjugate is composed of small latex microspheres, large latex microspheres, biotin, streptavidin, an adiponectin monoclonal antibody, an adiponectin polyclonal antibody and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride. After multiple research and development experiments, the large latex microspheres and the small latex microspheres are adopted; the large latex microspheres are coupled with the adiponectin monoclonal antibody, so that low-end sensitivity can be increased; the small latex microspheres are connected with the streptavidin and is mixed with the streptavidin proportionally; the adiponectin polyclonal antibody is connected with the biotin and is mixed with the biotin proportionally; and the two obtained mixtures are mixed proportionally and react with each other, and therefore, a linear rangecan be expanded; the two kinds of latex are mixed proportionally, so that repeatability and sensitivity can be improved, and a linear range can reach 1.00-32.00 mg/L.

Description

technical field [0001] The invention relates to the field of biotechnology, and relates to an adiponectin-latex enhanced immune turbidimetric kit, in particular to a preparation method of the adiponectin-latex enhanced immune turbidimetric kit. Background technique [0002] Adiponectin (APN), also known as Acrp30, GBP28 or AdipoQ, is a cytokine synthesized and secreted by adipocytes, which can exist stably in human plasma, with a concentration range of about 3.0-30.0 mg / L, accounting for about 0.01% of protein. Human adiponectin contains 244 amino acids, including 3 domains: an amino-terminal signal sequence, a collagen domain, and a carboxy-terminal globular domain. Adiponectin exists in blood in three isoforms: trimer (65kb), hexamer (150kb) and high molecular weight polymer (18-36 multimers, >280kb). Clinical studies have shown that adiponectin is closely related to obesity, type II diabetes, coronary heart disease, and insulin resistance, and has properties of anti-...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/531G01N33/53
CPCG01N33/531G01N33/53
Inventor 袁嘉扬单以朗
Owner 苏州普瑞斯生物科技有限公司
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