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Chemoluminescent immunoassay kit of hyaluronic acid and preparation method thereof

A technology of uronic acid chemistry and hyaluronic acid, which is applied in the field of immunoassay medicine, can solve the problems of low sensitivity, poor precision, pollution hazards of radioactive waste and other problems of enzyme immunoassay reagents, and achieves the effect of ensuring sensitivity and improving sensitivity.

Inactive Publication Date: 2009-09-16
CHEMCLIN DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to solve the short validity period of clinical radioimmunoassay reagents, the hazards of radioactive waste pollution, and the low sensitivity and poor precision of enzyme immunoassay reagents.

Method used

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  • Chemoluminescent immunoassay kit of hyaluronic acid and preparation method thereof
  • Chemoluminescent immunoassay kit of hyaluronic acid and preparation method thereof
  • Chemoluminescent immunoassay kit of hyaluronic acid and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0029] Example 1 Preparation of Hyaluronic Acid Chemiluminescence Immunoassay Assay Kit of the present invention

[0030] 1. Preparation of enzyme-labeled hyaluronic acid-binding protein and biotin-conjugated antibody

[0031] 1. The hyaluronic acid-binding protein was coupled to horseradish peroxidase by the glutaraldehyde method or the improved sodium periodate method.

[0032] 2. Preparation of biotin-conjugated hyaluronic acid polyclonal antibody or monoclonal antibody

[0033] (1) Dissolve biotin (BNHS) in N, N-dimethylformamide (DMF) by conventional methods to make 1 mg / mL;

[0034] (2) Use 0.1mol / L NaHCO with a pH value of 9.0 3 Dilute the purified hyaluronic acid polyclonal antibody or monoclonal antibody to 1-2 mg / mL;

[0035] (3) Mix according to the volume ratio of BNHS and antibody is about 1:8 or the weight ratio is about 1:7, and react for 2-4 hours under stirring at room temperature;

[0036] (4) Put it into 0.05 mol / L PBS with a pH value of 7.2, dialyze ove...

Embodiment 2-3

[0090] Example 2-3 Preparation of Hyaluronic Acid Chemiluminescence Immunoassay Assay Kit of the present invention

[0091] The hyaluronic acid chemiluminescent immunoassay assay kit was prepared in the same manner as in Example 1, except that plastic beads and plastic tubes were used as carriers.

Embodiment 4

[0092] Example 4 Preparation of Hyaluronic Acid Chemiluminescence Immunoassay Assay Kit of the present invention

[0093] Except that the magnetic particles were used as the carrier and the magnetic particle immobilized avidin was prepared by the EDC coupling method, the hyaluronic acid chemiluminescence immunoassay assay kit was prepared in the same manner as in Example 1.

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Abstract

The invention relates to the field of immunoassay medical science, and in particular provides a chemoluminescent immunoassay kit of a hyaluronic acid and a preparation method thereof. The kit comprises: 1) calibrating products of the hyaluronic acid; 2) an avidinylation solid phase carrier; 3) a biotinylation polyclonal antibody or monoclonal antibody of the hyaluronic acid; 4) a label of a hyaluronic acid binding protein; 5) a chemoluminescent substrate; and 6) a washing solution. Furthermore, the method for preparing the kit comprises the following steps: 1) preparing the calibrating objects from hyaluronic acid pure products ; 2) dermatating the carrier by an avidin; 3) performing biotinylation on the polyclonal antibody or monoclonal antibody of the hyaluronic acid; 4) labeling the hyaluronic acid binding protein by the label; 5) preparing the chemoluminescent substrate; 6) preparing the washing solution; 7) sub-packaging the calibrating products, the label, the chemoluminescent substrate and the washing solution; and 8) assembling the substances into the finished products. The kit has the advantages of simpleness, convenience, quickness, sensitiveness, stability, and the like.

Description

technical field [0001] The present invention relates to the field of immunoanalysis medicine, specifically, the present invention provides a hyaluronic acid (hyaluronic acid, hereinafter referred to as HA) biotin-avidin indirect coating technology combined with chemiluminescence immunoassay technology assay kit and its preparation method. Background technique [0002] Hyaluronic acid is a macromolecular amino polysaccharide synthesized by mesenchymal cells, participates in the formation of matrix in vivo, and is mainly taken up and decomposed by hepatic sinusoidal endothelial cells. Since HA is mainly metabolized in the liver, patients with liver diseases, especially those with liver cirrhosis, have HA metabolism disorders, and the concentration of HA in blood increases significantly, so HA is a good serological indicator of liver cirrhosis. The laboratory detection method of HA is mainly immunoassay, and the commonly used detection methods are radioimmunoassay (RIA) and en...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/66G01N21/76G01N33/577G01N33/543
Inventor 彭京胜应希堂宋胜利胡国茂郑金来
Owner CHEMCLIN DIAGNOSTICS CO LTD
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