Composition for detecting C peptide, application of composition, magnetic microsphere electrochemiluminescence immunoassay kit and detection method

A technology of luminescence immunity and composition, applied in the field of electrochemical detection, can solve the problems of low C-peptide value and inability to detect

Pending Publication Date: 2021-05-18
BEIJING UNIDIAG TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In such cases, insulin levels are high but C-peptide values ​​are low or undetectable

Method used

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  • Composition for detecting C peptide, application of composition, magnetic microsphere electrochemiluminescence immunoassay kit and detection method
  • Composition for detecting C peptide, application of composition, magnetic microsphere electrochemiluminescence immunoassay kit and detection method
  • Composition for detecting C peptide, application of composition, magnetic microsphere electrochemiluminescence immunoassay kit and detection method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Embodiment 1: the preparation of biotin-labeled C-peptide (C-Peptide) antibody and reagent Ra

[0054] The C-peptide antibody used to label biotin was purchased from Beijing Yuantian Xinye Technology Co., Ltd., the product number is YT-C-Peptide-002, and the clone number is 6E7.

[0055] Take 2.0 mg of the antibody used to label biotin C-peptide (C-Peptide), use the desalting column PD10 to replace the buffer with phosphate buffer (pH=7.8), use an ultrafiltration tube to concentrate and adjust the concentration to 2.0 mg / mL, Add 80 μg of biotin (dissolved in DMF), mix and react for 30 minutes, and use desalting column PD10 to remove unlabeled biotin. The biotin-labeled C-peptide (C-Peptide) antibody was diluted to 1 mg / L with 1% bovine serum albumin-containing phosphate buffer (pH=7.4) to serve as C-Peptide reagent Ra. The amount of biotin molecular labeling on the surface of each antibody molecule is 2-3.

Embodiment 2

[0056] Embodiment 2: Preparation of ruthenium-labeled C-peptide (C-Peptide) antibody and reagent Rb

[0057] The C-Peptide antibody used to label biotin was purchased from Beijing Yuantian Xinye Technology Co., Ltd., the article number is YT-C-Peptide-003, and the clone number is 4H3.

[0058] Take 2.0 mg of the C-Peptide antibody used to label ruthenium terpyridine, use the desalting column PD10 to replace the buffer with phosphate buffer (pH=7.8), use the ultrafiltration tube to concentrate and adjust the concentration to 2.0 mg / mL, add 80 μg of succinamide terpyridine ruthenium (dissolved in DMF), mix and react for 30 minutes, and use desalting column PD10 to remove unlabeled ruthenium. The ruthenium-labeled C-peptide (C-Peptide) antibody was diluted to 1 mg / L with phosphate buffer (pH=7.4) containing 1% bovine serum albumin, and used as C-Peptide reagent Rb. The number of ruthenium molecular labels on the surface of each antibody molecule is 5-6.

Embodiment 3

[0059] Embodiment 3: the preparation of calibrator

[0060] The antigen used for the preparation of the calibrator was purchased from Beijing Yuantian Xinye Technology Co., Ltd., and the product number is YT-C-Peptide-001. For recombinantly expressed proteins.

[0061] Use 1% bovine serum albumin-containing phosphate buffer (pH=7.4) to dilute the antigen to 0.025nmol / L and 5.400nmol / L according to the marked concentration. Used as a calibrator to establish a standard curve.

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Abstract

The invention relates to the field of electrochemical detection, and particularly relates to a composition for detecting C-Peptide, an application of the composition, a magnetic microsphere electrochemical luminescence immunoassay kit and a detection method. The method adopted by the invention is an electrochemical luminescence method, and ruthenium pyridine is adopted as a chemiluminescence marker so that the method has the obvious advantages that the sensitivity is better, the stability is better, ruthenium is a metal ion, the molecular weight is small, and the steric hindrance of an antibody is not influenced. The production process is short, the repeatability is good, and the detection range is wide. An electrochemical luminescence reaction is controllable, and the signal acquisition difficulty is reduced.

Description

technical field [0001] The present invention relates to the field of electrochemical detection, in particular to a composition for C-peptide (C-Peptide) detection, the application of the composition, a magnetic microsphere electrochemiluminescence immunoassay kit comprising the composition, and a composition based on the composition Magnetic microsphere electrochemiluminescence immunoassay method for object or kit. Background technique [0002] C-peptide is a polypeptide. It is part of the proinsulin molecule and has the following structure: β chain-Arg-Arg-C peptide-Lys-Arg-A chain. It is used to measure the secretory function of β-cell residues in diabetic patients receiving insulin therapy, so as to decide whether to switch to other anti-diabetic drugs or reduce the dosage of insulin. Diagnosis of endogenous hyperinsulinemia requires combined application of inhibition tests. Monitoring of patients after total pancreatectomy. It was suspected that the patient had secre...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/74G01N33/577G01N33/543G01N33/532G01N21/76
CPCG01N33/74G01N33/577G01N33/54326G01N33/532G01N21/76G01N2333/62
Inventor 秦军谢元东谢良思侯雪恬唐磊
Owner BEIJING UNIDIAG TECH
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