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Chemiluminescence immunoassay method for detecting S100 based on gold magnetic particles

A technology of chemiluminescent immunity and gold magnetic particles, which is applied in biological testing, material inspection products, etc., can solve the problems of antigen or antibody inactivation, complex coupling process, and restrictions on popularization and application, achieving high precision and good specificity , Operational safety effect

Inactive Publication Date: 2015-07-15
XIAN GOLDMAG NANOBIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The coupling process is complicated, and it is easy to inactivate the antigen or antibody during coupling. Not only the coupling efficiency is low, but also the cost is wasted. For these reasons, the popularization and application of chemiluminescence based on magnetic particles is limited to a certain extent.

Method used

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  • Chemiluminescence immunoassay method for detecting S100 based on gold magnetic particles
  • Chemiluminescence immunoassay method for detecting S100 based on gold magnetic particles
  • Chemiluminescence immunoassay method for detecting S100 based on gold magnetic particles

Examples

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Effect test

Embodiment 1

[0079] Example 1: Method for Quantitative Detection of S100 in Human Serum by Chemiluminescence Immunology Based on Gold Magnetic Particles

[0080] (1) Coated

[0081] (1.1) Pretreatment: Take 100 μl of 10 mg / ml gold magnetic particles, wash them twice with 100 μl of pH 7.4, 0.02M Tris-HCl equilibrium buffer to balance the pH of the magnetic particles.

[0082] (1.2) Coupling: Dissolve 160 μg S100 antibody in 200 μl pH7.4, 0.02M Tris-HCl coupling buffer, mix well and add to pretreated gold magnetic particles, shake at 37°C and 180rpm React in bed for 35min. After the reaction is complete, take it out, magnetically separate it for 2 minutes, and discard the supernatant.

[0083] (1.3) Washing: add 200 μl pH7.4, 0.02M Tris-HCl washing buffer containing 0.05% Tween-20, perform magnetic separation for 2 min, and discard the supernatant.

[0084] (2) Sealing: add 1ml containing 5% bovine serum albumin, 2.5% skimmed milk powder and 3% fetal bovine serum pH8.0, in the 0.01M PB bu...

Embodiment 2

[0089] Example 2: Method for Quantitative Detection of S100 in Human Serum by Chemiluminescence Immunology Based on Gold Magnetic Particles

[0090] (1) Coated

[0091] (1.1) Pretreatment: Take 100 μl of 10 mg / ml gold magnetic particles, wash them twice with 100 μl of pH 7.6, 0.02M Tris-HCl equilibrium buffer to balance the pH of the magnetic particles.

[0092] (1.2) Coupling: Dissolve 150 μg S100 antibody in 200 μl pH7.6, 0.02M Tris-HCl coupling buffer, mix well and add to pretreated gold magnetic particles, shake at 35°C, 200rpm The bed reacted for 38min. After the reaction is complete, take it out, magnetically separate it for 2 minutes, and discard the supernatant.

[0093] (1.3) Washing: Add 200 μl pH7.6, 0.02M Tris-HCl washing buffer containing 0.07% Tween-20, magnetically separate for 2 min, and discard the supernatant.

[0094] (2) Sealing: add 1ml containing 5% bovine serum albumin, 2.5% skimmed milk powder and 3% fetal bovine serum pH8.0, in the 0.01M PB buffer s...

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Abstract

The invention provides a chemiluminescence immunoassay method for detecting S100 based on gold magnetic particles. The method mainly comprises the following steps: (1) coating, namely taking gold magnetic particles as a carrier of immunoreaction and solid-phase separation, and coupling an S100 antibody onto the surface of the gold magnetic particles; (2) confining, namely confining the blank site at which the surface of the gold magnetic particles and the S100 antibody are bound by using confining liquid, performing magnetic separation, and removing the supernatant; (3) adding a to-be-detected sample and an enzyme-labeled secondary antibody which can be subjected to specific binding with an antigen into the gold magnetic particles which are confined in the step (2) and are bound with the S100 antibody for reacting, and forming a double-antibody sandwich complex; (4) cleaning; and (5) detecting. The method disclosed by the invention is high in detection sensitivity, high in specificity, wide in linearity range, high in precision, high in stability, safe to operate, simple and rapid, and radioactive contamination is avoided.

Description

technical field [0001] The invention relates to a method for chemiluminescent immunological detection of S100. Background technique [0002] The immunological detection methods for S100 protein currently on the market mainly include enzyme-linked immunoassay, electrochemiluminescence, radioimmunoassay, immunoradiometric assay, and fluorescent immunoassay. Enzyme-linked immunoassay can be used for quantitative detection, but the operation steps are cumbersome, greatly affected by human factors, and the sensitivity, specificity and application range vary greatly. Electrochemiluminescence quantitative detection has high accuracy and has the advantages of quantitative and stable levels, but it is not suitable for modern automatic immune instruments, and requires special supporting equipment. The investment cost is high, so it is difficult to popularize and apply. Radioimmunoassay and immunoradiometric assays have radioactive pollution and hazards, short half-lives of nuclides, ...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/68
Inventor 马乐张小梅吴松迪崔亚丽
Owner XIAN GOLDMAG NANOBIOTECH
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