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Kit for assaying anti-cyclic citrullinated peptide antibody and application thereof

An anti-cyclic citrullinated peptide and citrullinated peptide technology, which can be used in biological testing, general structural detail detection of gas analyzers, measuring devices, etc., and can solve the problems of improved detection sensitivity, long detection time, and low negative coincidence rate. problem, to achieve the effect of improving sensitivity, saving labor costs, and good stability

Active Publication Date: 2018-06-22
SUZHOU HAOOUBO BIOPHARML
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The third-generation CCP antigen is still a cyclic citrullinated peptide, but an epitope that cannot be recognized by Anti-CCP2 is added to it, and the detection sensitivity is slightly improved (73%)
However, the detection time of this kit is longer, it takes 2 hours, and the negative coincidence rate is low

Method used

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  • Kit for assaying anti-cyclic citrullinated peptide antibody and application thereof
  • Kit for assaying anti-cyclic citrullinated peptide antibody and application thereof
  • Kit for assaying anti-cyclic citrullinated peptide antibody and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1: Preparation of biotinylated antigen

[0049] Synthesis of citrulline 16 branched peptide:

[0050] According to the structural characteristics of citrulline and arginine, bioinformatics-related software is used to predict its spatial structure changes, and the known citrullined antigens are analyzed, classified, and screened out peptide libraries containing citrulline, which will contain citrulline A group of citrulline peptides was obtained by screening the polypeptide library of acid and rheumatoid arthritis serum, and four citrulline peptides with the highest combined detection sensitivity were screened. Using the characteristics of two amino groups of lysine, each citrulline peptide was citrulline peptide chains are coupled to form a molecule with four identical branched peptide chains, and finally four citrulline tetrabranched peptides are obtained, and then these four different branched peptide molecules are coupled into a molecule with lysine, which is...

Embodiment 2

[0065] Example 2: Preparation of alkaline phosphatase-labeled anti-human IgG antibody:

[0066] Materials and Instruments:

[0067] Materials: anti-human IgG antibody, alkaline phosphatase, coupling agent 2-iminosulfane hydrochloride, glycine, tris buffer.

[0068] Instruments: G-25 gel column, reagent low temperature storage box, Supperdex200 gel purification column, analytical balance, biochemical incubator;

[0069] Steps:

[0070] Step 1: Add 3mg of anti-human IgG antibody to 40mL of 2-iminosulfane hydrochloride coupling agent with a concentration of 10mg / mL, and let stand at 20°C for 20min;

[0071] Step 2: Add 2mL of 0.08mol / L glycine solution, let it stand at 20°C for 4min, use G-25 gel column to desalt, collect the activated anti-human IgG antibody, and store it at 5°C for later use;

[0072] Step 3: Add 3 mg alkaline phosphatase solution to 4 mg / mL 4-(N-maleimidomethyl)cyclohexane-1-carboxylic acid succinimide ester solution at 25°C Let it stand for 30 minutes, us...

Embodiment 3

[0075] Example 3: Preparation of calibrator: used for calibration curve.

[0076] 1. Materials and instruments:

[0077] Materials: anti-citrullinated peptide antibody, phosphate buffer saline, standard;

[0078] 2. Preparation steps:

[0079] Select an anti-citrullinated peptide antibody, dilute it in a certain ratio with a phosphate buffer solution with a pH of 7.5 and a substance concentration of 0.01 mol / L, and prepare 20RU / ml and 200RU / ml with reference to the standard. calibrator.

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Abstract

The invention relates to a kit for assaying an anti-cyclic citrullinated peptide antibody. The kit comprises a biotinylated antigen, wherein the biotinylated antigen is formed by coupling a pluralityof branched peptide units which have branched peptide chains through lysine, and each branched peptide unit is formed by coupling a plurality of citrullinated peptides through lysine. The kit for assaying the anti-cyclic citrullinated peptide antibody has the advantages that the detection sensitivity is high, the specificity is good, citrulline serves as one-component antigen after coupling, so that compared with a mixed antigen, the complexity of antigen component is reduced, the difficulty of quality control of a reagent is greatly reduced, the batch consistency is improved, quantitative detection can be achieved, and the time for completing all the processes is 45 minutes.

Description

technical field [0001] The invention belongs to the technical field of magnetic particle chemiluminescence immunodiagnosis, and in particular relates to a kit for measuring anti-cyclic citrullinated peptide antibody and its application. Background technique [0002] Rheumatoid arthritis (Rheumatoid Arthritis, RA) is the most common autoimmune disease and the most common chronic inflammatory joint disease, with a global incidence of about 1% and a Chinese incidence of 0.32-0.36%. RA is more common in women, with a male to female ratio of about 1:2-1:4. It is a systemic disease characterized by inflammation of the synovium of the joints, which progresses symmetrically from small to large joints, eventually leading to joint damage with soft tissue damage in later stages of the disease. [0003] Autoantibodies with higher specificity to RA include: rheumatoid factor (RF), anti-keratin antibody (AKA), anti-perinuclear factor (APF), RA33, and anti-cyclic citrullinated peptide ant...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/6854G01N33/6893G01N2800/102G01N33/564G01N2440/18
Inventor 崔利歌柳乐杨苏清赵婷郅晓乐黎静雯徐乐李裕明李庆春
Owner SUZHOU HAOOUBO BIOPHARML
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