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A kind of human s100 protein detection reagent and preparation method

A protein detection and reagent technology, which is applied in the field of laboratory medicine, can solve problems such as inconvenient development in conventional laboratories, patient costs, and increased patient burden, and achieve the effects of improving detection work efficiency, rapid detection, and easy operation

Active Publication Date: 2016-01-06
江苏美德医药技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method is divided into two types, one is the direct labeling method, which belongs to the instant luminous type, it is difficult to guarantee the stability and repeatability of the test results, and it requires special testing instruments
Not convenient for routine laboratory development
The second biotin-avidin-labeled electrochemiluminescence immunoassay method needs to be equipped with an expensive automatic electrochemiluminescence detector, which cannot be carried out in conventional laboratories, and it brings unnecessary costs to patients and increases the burden on patients

Method used

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  • A kind of human s100 protein detection reagent and preparation method
  • A kind of human s100 protein detection reagent and preparation method
  • A kind of human s100 protein detection reagent and preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] 1. The R1 reagent: HEPES buffer

[0066]

[0067]

[0068] After dissolving the above raw materials with 800ml of purified water, add purified water to make the volume to 1L, and then adjust the pH to 8.0 with hydrochloric acid or sodium hydroxide.

[0069] 2. The R2 reagent:

[0070] Step 1: Get 300nm carboxyl latex microspheres and dilute them into a suspension of 0.01mg / ml with 50mmol / L, PH6.0 MES buffer; add 20mgEDAC and 40mg of NHS to the carboxyl latex microsphere suspension in 1ml, Add EDAC and NHS and mix immediately, then react the mixture at room temperature for 15-30 minutes, stirring constantly;

[0071] Step 2: Wash the carboxyl latex microspheres with 50mmol / L, PH6.0 MES buffer or purified water to remove unreacted NHS and EDAC, and suspend the latex microspheres in purified water to obtain a concentration of 0.01mg / ml Carboxyl latex microsphere suspension;

[0072] Step 3: dissolving the mouse anti-human S100 antibody in 50 mmol / L, pH 8.5 TAP sal...

Embodiment 2

[0083] 1. The R1 reagent: Tris buffer

[0084]

[0085] After dissolving the above raw materials with 800ml of purified water, add purified water to make the volume to 1L, and then adjust the pH to 7.4 with hydrochloric acid or sodium hydroxide.

[0086] 2. The R2 reagent:

[0087] Step 1: Get 100nm carboxyl latex microspheres and dilute them into a suspension of 0.01mg / ml with 50mmol / L, PH6.0 MES buffer; add 20mgEDAC and 40mg of NHS to the carboxyl latex microsphere suspension in 1ml, Add EDAC and NHS and mix immediately, then react the mixture at room temperature for 15-30 minutes, stirring constantly;

[0088] Step 2: Wash the carboxyl latex microspheres with 50mmol / L, PH6.0 MES buffer or purified water to remove unreacted NHS and EDAC, and suspend the latex microspheres in purified water to obtain a concentration of 0.01mg / ml Carboxyl latex microsphere suspension;

[0089] Step 3: dissolving the mouse anti-human S100 antibody in 50 mmol / L, pH 8.5 TAP salt buffer solu...

Embodiment 3

[0099] 1. The R1 reagent: glycine buffer

[0100]

[0101] After dissolving the above raw materials with 800ml of purified water, add purified water to make the volume to 1L, and then adjust the pH to 8.0 with hydrochloric acid or sodium hydroxide. Among them, Thesit is polyoxyethylene lauryl ether, which belongs to the same category as Tween-20.

[0102] 2. The R2 reagent:

[0103] Step 1: Get 200nm carboxyl latex microspheres and dilute them into a suspension of 0.01mg / ml with 50mmol / L, PH6.0 MES buffer; add 20mgEDAC and 40mg of NHS to the carboxyl latex microsphere suspension in 1ml, Add EDAC and NHS and mix immediately, then react the mixture at room temperature for 15-30 minutes, stirring constantly;

[0104] Step 2: Wash the carboxyl latex microspheres with 50mmol / L, PH6.0 MES buffer or purified water to remove unreacted NHS and EDAC, and suspend the latex microspheres in purified water to obtain a concentration of 0.01mg / ml Carboxyl latex microsphere suspension; ...

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Abstract

The invention relates to a human S100 protein detection reagent and a preparation method thereof, and particularly relates to a reagent for detecting a human S100 protein by virtue of latex-enhanced immunoturbidimetry and a preparation method thereof. The reagent comprises a reagent R1, a reagent R2 and a standard product and is characterized in that the reagent R1 is a modified buffer solution with the pH value of 7.0-8.0; the reagent R2 is an anti-human S100 protein antibody latex reagent; the standard product is recombinant protein containing quantitative S100 protein or natural S100 protein standard liquid extracted from human serum. Compared with the prior art, the reagent provided by the invention has the characteristics of (1) high accuracy (the correlation R2 with the chemiluminescence immunoassay is 0.9956-0.9995) and (2) fast detection (only 10 minutes are needed from the test start to the result acquisition). Large-batch sample detection can be performed on a conventional biochemical analyzer, and the detection efficiency is greatly improved. The preparation method of the reagent is simple to operate, the raw materials are easily available, the cost is not high, and the method is suitable for various medical research institutions and conventional laboratories.

Description

technical field [0001] The invention relates to a reagent for detecting human S100 protein and a preparation method thereof, in particular to a reagent for detecting human S100 protein by latex-enhanced immune turbidimetry and a preparation method thereof, which can be applied in the field of laboratory medicine. Background technique [0002] Human central nervous system protein (S-100 protein) is mainly concentrated in the astrocytes of the central nervous system and corresponding tumor cells. The S-100 molecule is composed of α and β subunits, with a molecular weight of 21,000 and a biological half-life of 22 hours. . The S-100 protein in cerebrospinal fluid is related to age and increases with age, but the concentration of S-100 protein in plasma has no exact relationship with age and sex. When the central nervous system cells are damaged, the S100 protein seeps out from the cytosol into the cerebrospinal fluid, and then enters the blood through the damaged brain-blood b...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/566
CPCG01N33/531G01N33/538G01N33/6896G01N33/96G01N2496/00G01N2800/28
Inventor 陆上苏
Owner 江苏美德医药技术有限公司
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