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III type precollagen N end peptide chemiluminescence immune analysis quantitative determination reagent kit and preparing method thereof

A chemiluminescence immunity and chemiluminescence technology, which is applied in the field of immunoanalysis medicine, can solve the problems of inability to be widely used in clinical diagnosis and scientific research work, needs, and restrictions on promotion and use, so as 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

On the other hand, the chemiluminescence immunoassay kits in the prior art are all closed automatic chemiluminescence measurement systems, which require expensive automatic chemiluminescence measuring instruments, thus limiting the promotion and use, and cannot be widely used in clinical diagnosis and treatment. scientific research

Method used

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  • III type precollagen N end peptide chemiluminescence immune analysis quantitative determination reagent kit and preparing method thereof
  • III type precollagen N end peptide chemiluminescence immune analysis quantitative determination reagent kit and preparing method thereof
  • III type precollagen N end peptide chemiluminescence immune analysis quantitative determination reagent kit and preparing method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Preparation of the type III procollagen N-terminal peptide quantitative assay kit of the present invention

[0030] 1. Preparation of enzyme-labeled antibody

[0031] Type III procollagen N-terminal peptide monoclonal antibody was coupled with alkaline phosphatase by the glutaraldehyde method, fully dialyzed against PBS, added an equal volume of glycerol, and stored below -20°C.

[0032] 2. Preparation of type III procollagen N-terminal peptide calibrator

[0033] Prepared with pure N-terminal peptide of type III procollagen, divided into 6 bottles of 0, 6.2, 12.5, 25, 50, 100ng / ml.

[0034] 3. Enzyme-labeled antibody concentration selection

[0035] Use the square array method to select the working concentration of the enzyme-labeled antibody to be greater than 1:6000.

[0036] 4. Preparation of solid-phase coated plates

[0037] (1) Coated

[0038] Trisodium citrate 2.92g

[0039] Citric acid 1.78g

[0040] Double distilled water 400ml

[0041] After...

Embodiment 2～3

[0071] Examples 2-3 Preparation of the Type III procollagen N-terminal peptide quantitative assay kit of the present invention

[0072] Divided by horseradish peroxidase-labeled type III procollagen N-terminal peptide monoclonal antibody, with luminol (luminol) system as chemiluminescent substrate, this chemiluminescent substrate includes A liquid and B liquid, based on 1000mL of the chemical Luminescence substrate A solution, including 1.7716g luminol, 0.051g 4-hydroxybiphenyl, 0.012g 4-iodophenylboronic acid, 11.4g boric acid, 4.9g borax, its pH value is 8.0-10.0; based on 1000mL of the chemical Luminescent substrate solution B, including 0.329g carbamide peroxide, 1ml Tween20, 51.58g Na 2 HPO 4 12H 2 O, 8.74gNaH 2 PO 4 2H 2 O, its pH value is 7.0 ~ 7.6, with plastic beads and plastic tubes as solid phase carriers respectively, except that the pH value of the coating solution is 4.5, and the pH value of the blocking solution is 7.5, the rest are all in the same way as i...

Embodiment 4

[0073] Example 4 Preparation of the type III procollagen N-terminal peptide quantitative assay kit of the present invention

[0074] Except that the magnetic particles were used as the carrier, and the magnetic particles were coated by the glutaraldehyde method, the rest were prepared in the same way as in Example 1 to prepare the type III procollagen N-terminal peptide quantitative assay kit.

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Abstract

The invention relates to the medical field of immunoassay, more specially, the invention provides a chemiluminescent immunoassay quantitative detection kit for III-type procollagen N-terminal peptide (PIIINP) and a preparation method thereof. The kit of the invention comprises 1) III-type procollagen N-terminal peptide calibrators, 2) solid-phase vectors which are coated with III-type procollagen N-terminal peptide monoclonal antibodies, 3) enzyme markers for III-type procollagen N-terminal peptide monoclonal antibodies, 4) chemiluminescent substrates and 5) concentrated washing solution. Further, the preparation method of the kit according to the invention comprises the following steps: 1) preparing the III-type procollagen N-terminal peptide calibrators with pure III-type procollagen N-terminal peptide, 2) coating the vectors with III-type procollagen N-terminal peptide monoclonal antibodies, 3) marking the III-type procollagen N-terminal peptide monoclonal antibodies with the enzyme, 4) preparing the chemiluminescent substrates, 5) preparing the concentrated washing solution, 6) packaging the III-type procollagen N-terminal peptide calibrators, the enzyme markers, the chemiluminescent substrates and the concentrated washing solution and 7) assembling finished products. The kit of the invention has the advantages of simplicity, convenience, rapidness, sensitivity, stability and the like.

Description

technical field [0001] The invention relates to the field of immunoanalysis medicine. Specifically, the invention provides a type III procollagen N-terminal peptide chemiluminescent immunoassay quantitative assay kit and a preparation method thereof. Background technique [0002] Liver fibrosis is an inevitable stage in the development of various chronic liver diseases to liver cirrhosis. Liver cirrhosis is a malignant disease that seriously threatens human life and health. Early diagnosis and treatment of liver fibrosis is one of the effective means to fight against liver cirrhosis, therefore, early diagnosis of liver fibrosis is very important. [0003] Currently, serum markers commonly used in clinical diagnosis of liver fibrosis include type III procollagen N-terminal peptide, type IV collagen, type III procollagen N-terminal peptide, prolyl hydroxylase, laminin, and laminin-P1 Fragment, TGF-β1, etc., commonly used type III procollagen N-terminal peptide, type IV colla...

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 高鹏应希堂宋胜利胡国茂郑金来于尚永
Owner CHEMCLIN DIAGNOSTICS CO LTD
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