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Value setting method for C reactive proteins in serum

A technology for reacting protein and serum, which can be used in material separation, material inspection products, instruments, etc., and can solve the problems of low total protein yield, high price, time-consuming and labor-intensive, etc.

Active Publication Date: 2020-02-28
NAT INST OF METROLOGY CHINA
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the yield of isotope-labeled whole protein is low, time-consuming and labor-intensive, and the price is extremely expensive, and there are slight differences in the tertiary structure of natural proteins and their isotope-labeled analogs

Method used

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  • Value setting method for C reactive proteins in serum
  • Value setting method for C reactive proteins in serum
  • Value setting method for C reactive proteins in serum

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Embodiment Construction

[0037] Reagent:

[0038] Acetonitrile was purchased from German Merck company product;

[0039] Ultrapure water is purified by Millipore pure water system;

[0040] Trifluoroacetic acid was purchased from Sigma-Aldirich, USA;

[0041] Formic acid was purchased from Fisher Scientific, USA;

[0042] Dithiothreitol (DTT) was purchased from U.S. inalco company;

[0043]Trypsin was purchased from Promega, USA;

[0044] Tris, urea (Urea) were purchased from American Amresco company;

[0045] Iodoacetamide (Iodoacetamide, IAM) was purchased from U.S. Inalco Company;

[0046] Magnetic beads were purchased from Invitrogen;

[0047] EK, GK, RK, L-EK, L-GK, and L-RK were synthesized by Nanjing GenScript Biotechnology Co., Ltd. (MALDI-TOF-MS was used to analyze the digested CRP, and the mass spectrometry response was high, and the CRP that could be separated was selected. The three peptides EK, GK, and RK; L-EK, L-GK, and L-RK are isotope-labeled peptides of these three peptides). ...

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Abstract

The invention discloses a value setting method for C reactive proteins in serum. The method comprises the following steps of (1) purification and enrichment of C reactive proteins (CRP)in serum; (2) purity detection of the C reactive proteins; (3) digestion; and (4) quantitative analysis of the C reactive proteins. The method does not need to consider the factors such as protein extraction rate, digestion efficiency and instrument stability, and only needs to consider the correctness of pure CRP concentration in the whole experiment; the used pure CRP solution is a national primary standard substance (developed by the National Institute of Measurement) which is reliable in numerical value and can be traced to an SI unit; and the raw material is nature CRP which does not need to consider the differences of structures and is more excellent than the isotope labeled whole proteins.

Description

technical field [0001] The invention relates to the technical field of protein content determination, in particular to a method for determining the value of C-reactive protein in serum. Background technique [0002] C-reactive protein is a pentameric serum protein released by the liver, which can be used to monitor cardiovascular disease and systemic inflammation, and is one of the important clinical markers. Because of its good stability and the commercialization of monoclonal antibodies, it has clinical application value. The commonly used detection methods include enzyme-linked immunosorbent assay, immune projection turbidimetry, and immunoluminescence. However, due to the development of methods and reagents for different targets by various manufacturers, the measurement deviation is very large. Therefore, it is urgent to develop a reference measurement method for CRP. [0003] Isotope dilution mass spectrometry has the characteristics of high sensitivity, high quantita...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/577G01N33/543G01N27/447G01N30/72
CPCG01N27/447G01N33/54326G01N33/577G01N33/6848G01N33/6893G01N2333/4737G01N2800/32G01N2800/7095
Inventor 宋德伟朱文陈鸿飞李红梅孙浩峰肖鹏刘建仪马凌云王馨雪
Owner NAT INST OF METROLOGY CHINA
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