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Electrical chemiluminescence immunoassay based on gold magnetic particles

A technology of luminescent immunity and gold magnetic particles, which is applied in the field of medical testing, can solve the problems of lack of versatility, poor controllability of the reaction, and cumbersome steps, and achieve the effects of low labeling cost, easy operation, and simplified experimental steps

Inactive Publication Date: 2012-10-10
XIAN GOLDMAG NANOBIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The invention provides an electrochemiluminescence immunoassay method based on gold magnetic particles, which can quantitatively detect the content of antigen / antibody in human serum, plasma, cell culture supernatant or other related biological fluids, so as to solve the problem of relatively low cost in the prior art. High, cumbersome steps, poor response controllability, no versatility, etc.

Method used

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  • Electrical chemiluminescence immunoassay based on gold magnetic particles
  • Electrical chemiluminescence immunoassay based on gold magnetic particles
  • Electrical chemiluminescence immunoassay based on gold magnetic particles

Examples

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Embodiment 1

[0035] Example 1: Quantitative detection of PSA concentration in human serum by electrochemiluminescence immunoassay method based on gold magnetic particles.

[0036] 1) Preparation of immunomagnetic beads with gold magnetic particles

[0037] 1.1) Prepare PSA monoclonal antibody (capture antibody) to 1mg / mL with coupling buffer, take out a certain amount of antibody solution in a centrifuge tube, and mark it as pre, and reserve it for coupling efficiency detection.

[0038] 1.2) Pipette with a pipette Put 200 μL of gold magnetic particles (5 mg / mL) in a 2 mL centrifuge tube, add 400 μL of coupling buffer and shake gently to mix, magnetically separate for 2 min, and discard the supernatant.

[0039] 1.3) Take 300 μL of this antibody solution and add it to the In the centrifuge tube of gold magnetic particles, shake gently to mix, fix the centrifuge tube on a constant temperature shaker, 37 ° C, 180rpm shaking reaction for 1h, after the reaction is completed, magnetically s...

Embodiment 2

[0067] Example 2: Quantitative detection of the concentration of AFP in human serum by electrochemiluminescence immunoassay method based on gold magnetic particles.

[0068] In addition to coating gold magnetic particles with AFP antibody (capture antibody) when preparing immunomagnetic beads, Ru(bpy) 3 2+ Labeled AFP antibody (detection antibody), Ru(bpy) 3 2+ Antibody and Ru(bpy) in the process of labeling AFP antibody 3 2+ -Except for the molar ratio of NHS ester and the concentration of several AFP standard substances selected in the process of establishing the standard curve, other steps are the same as in Example 1.

Embodiment 3

[0069] Example 3: Quantitative detection of TSH concentration in human serum by electrochemiluminescence immunoassay method based on gold magnetic particles.

[0070] In addition to coating gold magnetic particles with TSH antibody (capture antibody) when preparing immunomagnetic beads, Ru(bpy) 3 2+ Labeled TSH antibody (detection antibody), Ru(bpy) 3 2+ Antibody and Ru(bpy) in the process of labeling TSH antibody 3 2+ -Except for the molar ratio of NHS ester and the concentrations of several TSH standard substances selected during the establishment of the standard curve, other steps are the same as in Example 1.

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Abstract

The invention provides an electrical chemiluminescence immunoassay method based on gold magnetic particles, which realizes the quantitative detection on the content of antigen / antibody in human serum, blood plasma, cell culture supernatant or other related biological liquid so as to solve the problems of high cost, complicated steps, poor reaction controllability, no generality and the like in the prior art. The method mainly comprises the steps of 1) preparing immunomagnetic beads by gold magnetic particles; 2) labeling the antigen / antibody by Ru(bpy) <2+>3; 3) establishing a standard curve for quantitatively detecting the antigen / antibody; and 4) detecting a sample to be detected. A novel electrical chemiluminescence immunoassay technology is adopted, and the electrical chemiluminescence immunoassay method based on gold magnetic particles has the advantages that the operation is simple and convenient, the valence of antibody can not be affected in the process that the antibody is labeled by Ru(bpy) <2+>3, the labeling cost is low, and the universality is realized.

Description

technical field [0001] The invention belongs to the field of medical inspection, and in particular relates to electrochemiluminescence immunoassay technology. Background technique [0002] The concentration of a certain antigen / antibody in the human body may be directly related to its health status, physiological function, and physiological cycle, so quantitative detection of the concentration of antigen / antibody in the human body is of great significance to the human body. For example, the concentration of PSA in human serum is of great significance in the diagnosis, prognosis and curative effect detection of prostate cancer. At present, the detection techniques for antigens / antibodies in human serum, plasma, cell culture supernatant or other related biological fluids mainly include: Radioimmunoassay (RIA), Enzyme linked Immunosorbent Assay (ELISA), Fluorescence Immunoassay (FIA), Chemiluminescence Immunoassay (CLIA), Immunochromatography (Lateral Flow Immunoassay, LFIA) a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/551G01N33/531
Inventor 张秦鲁成晓崔亚丽
Owner XIAN GOLDMAG NANOBIOTECH
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