III type procollagen N-terminal peptide quantitative measurement kit and preparation method thereof

A technology for quantitative determination and procollagen, applied in the field of immunodiagnosis, can solve the problems of narrow detection range, poor sensitivity, and easy error of experimental results, and achieve the effect of expanding the reaction surface area, improving the precision of reagents, and being easy to operate.

Inactive Publication Date: 2016-10-26
威海威高生物科技有限公司
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  • Abstract
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  • Application Information

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Problems solved by technology

However, the enzyme-linked immunoassay and chemiluminescence method take a long time to detect, mainly rely on a series of tedious operations such as pure manual sample addition, and the efficiency is low, resulting in large errors in the experimental results, while the enzymatic reaction is not thorough enough and is easily affected by external interference factors, such as Temperature, time, and material concentration, etc., so the detection has low specificity, poor sensitivity, and narrow detection range
The advantages of radioimmunoassay are sensitivity, specificity, simplicity, and small sample volume, etc., but the half-life period of reagents is expensive, and special equipment such as scintillation counters are required, and radionuclides have certain potential hazards to the human body. Sexuality, difficult disposal of experimental waste, radionuclide labeling sometimes alters the physiological activity of certain biological substances

Method used

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  • III type procollagen N-terminal peptide quantitative measurement kit and preparation method thereof
  • III type procollagen N-terminal peptide quantitative measurement kit and preparation method thereof
  • III type procollagen N-terminal peptide quantitative measurement kit and preparation method thereof

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Embodiment 1

[0052] 1) Preparation of magnetic particle-labeled PⅢNP antibody

[0053] After mixing magnetic particles with carboxyl groups and EDC at a weight ratio of 1:1, 1:2, and 1:3, 15 μg, 20 μg, 25 μg, and 30 μg of PⅢNP monoclonal antibody were added to each mg of magnetic particles, respectively. React at 23°C and 26°C for 1 hour, add a certain amount of glycine to block the redundant sites of EDC, so that the concentration reaches 25mM, and react at 20°C, 23°C, and 26°C for 0.5 hours respectively. Store the labeled magnetic particles in 0.01M PBS containing 1% bovine serum albumin at 2-8°C;

[0054] 2) Preparation of isoluminol-labeled PⅢNP antibody

[0055] Add isoluminol to the PⅢNP antibody for labeling, the concentration of glutaraldehyde used is 0.75%, 1.0%, 1.25%, and 1.5%, respectively, and 0.05mg, 0.10mg, 0.15mg, 0.20 mg, the optimal labeling ratio of antibody to isoluminol is 10:1, react at 20°C, 23°C, and 26°C for 1.5 hours, dialyze with 0.01M PBS with pH 7.2-7.4, add ...

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Abstract

The invention provides a magnetic particle chemiluminescence method III type procollagen N-terminal peptide quantitative measurement kit and a preparation method thereof. The kit is composed of PIIINP magnetic particles, tracing conjugate for measuring PIIINP, and a quality control substance. The invention also provides a preparation method of the quantitative measurement kit. The preparation method adopts a micro particle chemiluminescence immunoassay technology, an automatic chemiluminescence method is used to carry out detection, the operation time is reduced, the artificial operation error is reduced, the detection precision and accuracy are improved, and the kit is suitable for assisted diagnosis of early stage hepatic fibrosis in clinic.

Description

technical field [0001] The invention belongs to the technical field of immunodiagnosis, and relates to a quantitative assay kit, in particular to a magnetic particle chemiluminescent immunoassay kit for measuring the N-terminal peptide of type III procollagen, and the invention also relates to a preparation method of the kit , and a method for using the kit to measure the concentration of type III procollagen N-terminal peptide. Background technique [0002] Liver fibrosis refers to the abnormal proliferation of connective tissue in the liver caused by various pathogenic factors, leading to the pathological process of excessive precipitation of diffuse extracellular matrix in the liver. It is not an independent disease, and many chronic liver diseases can cause liver fibrosis. The etiology can be roughly divided into infectious (chronic B, C and D viral hepatitis, schistosomiasis, etc.), congenital metabolic defects (hepatolenticular degeneration, hemochromatosis, α1-antitry...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/553G01N33/532G01N21/76
CPCG01N33/6893G01N21/763G01N33/532G01N33/54326G01N33/553
Inventor 蒙红胜乔文革吴晗琪王凌凌王文艳
Owner 威海威高生物科技有限公司
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