Laminin chemiluminescence immune analysis determination reagent kit and preparing method thereof

A chemiluminescence immunoassay and laminin technology, applied in the field of immunoassay medicine, can solve the problems of inability to widely apply clinical diagnosis and scientific research work, needs, and limited popularization and use, to ensure sensitivity, increase sensitivity, and facilitate operation and production. Effect

Inactive Publication Date: 2009-03-04
CHEMCLIN DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, the chemiluminescence immunoassay kit in the prior art is a closed fully automatic chemiluminescence measurement system, which requires an expensive fully automatic chemiluminescence measuring instrument, thus limiting its popularization and use, and cannot be widely used in clinical diagnosis and scientific research.

Method used

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  • Laminin chemiluminescence immune analysis determination reagent kit and preparing method thereof
  • Laminin chemiluminescence immune analysis determination reagent kit and preparing method thereof
  • Laminin chemiluminescence immune analysis determination reagent kit and preparing method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Preparation of the laminin chemiluminescence immunoassay assay kit of the present invention

[0038] 1. Preparation of enzyme-labeled antibody and preparation of biotin-conjugated antibody

[0039] 1. The polyclonal antibody or monoclonal antibody of laminin is coupled with horseradish peroxidase by periodic acid oxidation method, fully dialyzed against PBS, added an equal volume of glycerol, and stored below -20°C for use.

[0040] 2. Preparation of biotin-conjugated laminin monoclonal antibody

[0041] (1) Use conventional methods to dissolve biotin (BNHS) in N,N-dimethylformamide (DMF) to make 1mg / mL;

[0042] (2) Use 0.1mol / L NaHCO with a pH of 9.0 3 Dilute the purified laminin monoclonal antibody to 1~2mg / mL;

[0043] (3) Mix according to the volume ratio of BNHS and antibody of 1:8 or weight ratio of 1:7, and react for 2 to 4 hours under stirring at room temperature;

[0044] (4) Load the dialysis to 0.05mol / L PBS with a pH of 7.2, dialyze overnight at 4°C, ...

Embodiment 2～3

[0096] Examples 2 to 3 Preparation of the laminin chemiluminescence immunoassay assay kit of the present invention

[0097] Except that plastic beads and plastic tubes are used as carriers, the rest are prepared in the same manner as in Example 1 to prepare a laminin chemiluminescence immunoassay assay kit.

Embodiment 4

[0098] Example 4 Preparation of the laminin chemiluminescence immunoassay assay kit of the present invention

[0099] In addition to using the glutaraldehyde method to label the laminin monoclonal antibody with alkaline phosphatase, using AMPPD as the chemiluminescent substrate, using plastic beads as the carrier, the coating solution is a citrate buffer with a pH of 4.5, and a blocking solution. Except for the pH value of 7.5, the rest were prepared using the same method as in Example 1 to prepare a laminin chemiluminescence immunoassay assay kit.

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Abstract

The invention relates to the medical field of immunoassay, more specially, the invention provides a laminin detection kit and a preparation method thereof. The kit of the invention comprises 1) a laminin calibrator, 2) an avidin-coated vector, 3) a mixture of a biotinylation laminin monoclonal antibody and a laminin polyclonal antibody or a monoclonal antibody enzyme marker, 5) a chemiluminescent substrate and 6) a concentrated washing solution. Further, the preparation method of the kit according to the invention comprises the following steps: 2) preparing the calibrator with laminin, 2) coating the vector with avidin, 3) biotinylating the laminin monoclonal antibody, 4) marking the laminin polyclonal antibody or monoclonal antibody with the enzyme, 5) preparing the chemiluminescent substrate, 6) preparing the concentrated washing solution, 7) packaging the calibrator, the biotinylation laminin, the chemiluminescent sustrate and the concentrated washing solution, and 8) assembling finished products. The kit of the invention has the advantages of simplicity, convenience, sensitivity, stability and the like, and provides a valuable detection means for clinic diagnosis and scientific research.

Description

Technical field [0001] The present invention relates to the field of immunoassay medicine. Specifically, the present invention provides a laminin (LN) biotin-avidin immunoamplification technique combined with chemiluminescence immunoassay technique determination kit and a preparation method thereof. Background technique [0002] Liver fibrosis is the pre-pathological stage of liver cirrhosis. It is a wound healing process in which extracellular matrix is ​​reversibly deposited when the liver is chronically injured. Many chronic liver diseases can cause fibrosis. At present, the main diagnostic methods for liver fibrosis include imaging diagnosis, pathological diagnosis and serological diagnosis. Imaging diagnosis is widely used in clinical practice, but abnormal images can only appear when liver cirrhosis and portal hypertension occur in the late stage of liver fibrosis. The diagnostic gold index of liver fibrosis-liver biopsy, due to the small amount of specimens taken and the h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N21/76G01N33/577G01N33/543
Inventor 蒋冰飞应希堂胡国茂唐宝军张坤
Owner CHEMCLIN DIAGNOSTICS CO LTD
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