Kit for determining concentration of Anti-cyclic citrullinated peptide antibody and preparation method thereof

Abstract

The invention relates to the technical field of biology, in particular to a kit for determining concentration of Anti-cyclic citrullinated peptide antibody. The invention adopts the following technical scheme: the kit for determining concentration of Anti-cyclic citrullinated peptide antibody is characterized by comprising a reagent R1 and a reagent R2, wherein the reagent R1 comprises a buffer solution, a promoter, and a preservative, and the reagent R2 comprises a buffer solution, a stabilizing agent, latex microspheres conjugated with Cyclic citrullinated peptide antigens-biotin-streptavidin, and a preservative. The kit for determining concentration of Anti-cyclic citrullinated peptide antibody has the advantages that: according to the kit for determining concentration of Anti-cyclic citrullinated peptide antibody, the latex microspheres are connected with streptavidin at a ratio of 1: 1, the Cyclic citrullinated peptide antigens is connected with biotin at a ratio of 1: 2, and therepeatability and sensitivity can be obviously improved by a 1: 1 mixing reaction and the repeatability and the linear range can be from 5 to 150 U / ml.

Description

technical field

[0001] The invention relates to the field of biotechnology, in particular to a kit for measuring the concentration of anti-cyclic citrullinated peptide and a preparation method thereof. Background technique

[0002] Cyclic citrulline polypeptide antibody (anti-cyclic peptide containing citrulline, anti-CCP): It is a polypeptide fragment of cyclic polyguanidine, mainly IgG type antibody. Anti-CCP antibodies are spontaneously secreted by B lymphocytes of RA patients, while B lymphocytes of patients with other diseases and normal people do not spontaneously secrete anti-CCP antibodies. Therefore, anti-CCP antibodies have high specificity for RA.

[0003] Rheumatoid arthritis (RA) is a common systemic autoimmune disease, characterized by joint synovial inflammation and symmetrical and destructive joint lesions. big pain. The incidence rate of this disease in my country is about 0.4%, and it is 0.5-1% in the world, and the age of onset of patients is 20-50 year...

Examples

Embodiment 1

[0041] A kit for measuring the concentration of an anti-cyclic citrullinated peptide antibody, comprising reagent R1 and reagent R2, the components and concentrations of the reagent R1 include: as the first buffer A, sodium dihydrogen phosphate dihydrate, the concentration of which is 0.8g / L, as the first buffer solution B, disodium hydrogen phosphate dodecahydrate, its concentration is 24.31g / L, as the first electrolyte sodium chloride, its concentration is 5.8g / L, as the first highest PEG 6000 as a molecular accelerator at a concentration of 40 g / L, and Proclin-950 as a primary preservative at a concentration of 2.0%.

[0042] The reagent R2 includes a second buffer, a preservation solution, latex microspheres coupled with cyclic citrullinated peptide antigen-biotin-streptavidin, and the second buffer is 2-(N-morpholine ) ethanesulfonic acid monohydrate, the concentration is 8.05g / L; the preservation solution includes: N, N-dihydroxyethylglycine with a concentration of 9.83g...

Embodiment 2

[0054] A test kit for measuring the concentration of an anti-cyclic citrullinated peptide antibody, comprising reagent R1 and reagent R2, each component and concentration of the reagent R1 including sodium dihydrogen phosphate dihydrate as the first buffer solution A, the concentration of which is 3.2g / L, as the first buffer B, disodium hydrogen phosphate dodecahydrate, its concentration is 8.75g / L, as the first electrolyte sodium chloride, its concentration is 20.0g / L, as the first polymer Accelerator PEG 6000, its concentration is 80g / L, as the first preservative Proclin-300, its concentration is 0.8%, and the reagent R2 includes the second buffer solution, preservation solution, coupled with cyclic citrulline peptide The latex microspheres of antigen-biotin-streptavidin, the second buffer is 2-(N-morpholine) ethanesulfonic acid monohydrate, and its concentration is 27.30g / L; The preservation solution includes : Concentration is 5.0g / L of N,N-dihydroxyethylglycine, casein as...

Embodiment 3

[0066] A test kit for measuring the concentration of an anti-cyclic citrullinated peptide antibody, comprising reagent R1 and reagent R2, each component and concentration of the reagent R1 including sodium dihydrogen phosphate dihydrate as the first buffer A, the concentration of which is 2g / L, as the disodium hydrogen phosphate dodecahydrate of the first buffer solution B, its concentration is 16g / L, as the sodium chloride of the first electrolyte, its concentration is 12g / L, as the first polymer accelerator PEG 6000, its concentration is 120g / L, as the first preservative thimerosal, its concentration is 1.0%, and the reagent R2 includes the second buffer solution, preservation solution, coupled with cyclic citrullinated peptide antigen-biotin- For the latex microspheres of streptavidin, the second buffer is 2-(N-morpholine) ethanesulfonic acid monohydrate, and its concentration is 16.80g / L; the preservation solution includes: a concentration of 1.26g N,N-dihydroxyethylglycin...

technical field

[0001] The invention relates to the field of biotechnology, in particular to a kit for measuring the concentration of anti-cyclic citrullinated peptide and a preparation method thereof. Background technique

[0002] Cyclic citrulline polypeptide antibody (anti-cyclic peptide containing citrulline, anti-CCP): It is a polypeptide fragment of cyclic polyguanidine, mainly IgG type antibody. Anti-CCP antibodies are spontaneously secreted by B lymphocytes of RA patients, while B lymphocytes of patients with other diseases and normal people do not spontaneously secrete anti-CCP antibodies. Therefore, anti-CCP antibodies have high specificity for RA.

[0003] Rheumatoid arthritis (RA) is a common systemic autoimmune disease, characterized by joint synovial inflammation and symmetrical and destructive joint lesions. big pain. The incidence rate of this disease in my country is about 0.4%, and it is 0.5-1% in the world, and the age of onset of patients is 20-50 year...

Examples

Embodiment 1

[0041] A kit for measuring the concentration of an anti-cyclic citrullinated peptide antibody, comprising reagent R1 and reagent R2, the components and concentrations of the reagent R1 include: as the first buffer A, sodium dihydrogen phosphate dihydrate, the concentration of which is 0.8g / L, as the first buffer solution B, disodium hydrogen phosphate dodecahydrate, its concentration is 24.31g / L, as the first electrolyte sodium chloride, its concentration is 5.8g / L, as the first highest PEG 6000 as a molecular accelerator at a concentration of 40 g / L, and Proclin-950 as a primary preservative at a concentration of 2.0%.

[0042] The reagent R2 includes a second buffer, a preservation solution, latex microspheres coupled with cyclic citrullinated peptide antigen-biotin-streptavidin, and the second buffer is 2-(N-morpholine ) ethanesulfonic acid monohydrate, the concentration is 8.05g / L; the preservation solution includes: N, N-dihydroxyethylglycine with a concentration of 9.83g...

Embodiment 2

[0054] A test kit for measuring the concentration of an anti-cyclic citrullinated peptide antibody, comprising reagent R1 and reagent R2, each component and concentration of the reagent R1 including sodium dihydrogen phosphate dihydrate as the first buffer solution A, the concentration of which is 3.2g / L, as the first buffer B, disodium hydrogen phosphate dodecahydrate, its concentration is 8.75g / L, as the first electrolyte sodium chloride, its concentration is 20.0g / L, as the first polymer Accelerator PEG 6000, its concentration is 80g / L, as the first preservative Proclin-300, its concentration is 0.8%, and the reagent R2 includes the second buffer solution, preservation solution, coupled with cyclic citrulline peptide The latex microspheres of antigen-biotin-streptavidin, the second buffer is 2-(N-morpholine) ethanesulfonic acid monohydrate, and its concentration is 27.30g / L; The preservation solution includes : Concentration is 5.0g / L of N,N-dihydroxyethylglycine, casein as...

Embodiment 3

[0066] A test kit for measuring the concentration of an anti-cyclic citrullinated peptide antibody, comprising reagent R1 and reagent R2, each component and concentration of the reagent R1 including sodium dihydrogen phosphate dihydrate as the first buffer A, the concentration of which is 2g / L, as the disodium hydrogen phosphate dodecahydrate of the first buffer solution B, its concentration is 16g / L, as the sodium chloride of the first electrolyte, its concentration is 12g / L, as the first polymer accelerator PEG 6000, its concentration is 120g / L, as the first preservative thimerosal, its concentration is 1.0%, and the reagent R2 includes the second buffer solution, preservation solution, coupled with cyclic citrullinated peptide antigen-biotin- For the latex microspheres of streptavidin, the second buffer is 2-(N-morpholine) ethanesulfonic acid monohydrate, and its concentration is 16.80g / L; the preservation solution includes: a concentration of 1.26g N,N-dihydroxyethylglycin...

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