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Circulating immune complex of peroxide reductase IV and application of complex

An immune complex, peroxide technology, used in the medical field, can solve problems such as life-threatening, labor loss, and multi-organ involvement in the whole body

Active Publication Date: 2013-09-18
CHONGQING LIANBAI BOCHAO MEDICAL EQUIP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If RA is not effectively treated in the early stage, it will cause joint dysfunction and even loss of labor force, and lead to the involvement of multiple organs in the whole body, which will be life-threatening

Method used

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  • Circulating immune complex of peroxide reductase IV and application of complex
  • Circulating immune complex of peroxide reductase IV and application of complex
  • Circulating immune complex of peroxide reductase IV and application of complex

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0080] Example 1 Application of co-immunoprecipitation combined with two-dimensional gel electrophoresis to confirm the existence of circulating immune complexes of peroxide reductase IV in serum

[0081] The invention verifies whether there is circulating immune complex of peroxide reductase IV through immunoproteomics method, especially whether the immune complex of peroxide reductase IV exists in the circulation system of patients with rheumatoid arthritis.

[0082] One test principle

[0083] Immune complexes exist in the form of complexes of antigens and antibodies, and Protein G Sepharose4B has the property of binding to antibody IgG. Therefore, protein G Sepharose4B is first used to adsorb antibodies in serum, so that the immune complex is also captured by Protein G Sepharose4B along with antibodies; Valence cross-linking, so that the antibodies in the serum can be stably fixed on the Protein G Sepharose microspheres, and cannot be separated by the swelling solution, b...

Embodiment 2

[0094] Example 2 Application of Peroxide Reductase IV Circulating Immune Complex Positive Control to Confirm the Existence of the Circulating Immune Complex in Serum

[0095] 1. Generation of peroxide reductase IV immune complexes in vitro

[0096] 1. Obtain recombinantly expressed peroxide reductase IV and mouse anti-human peroxide reductase IV monoclonal antibody;

[0097] 2. According to the molar ratio of 3:1, mix peroxide reductase IV and mouse anti-human peroxide reductase IV monoclonal antibody at 4°C overnight to make the two form an immune complex;

[0098] 3. Centrifuge the overnight reaction solution at 14,000 rpm at 4°C for 15 minutes. Due to the large molecular weight of the immune complex, remove the supernatant, and the remaining precipitate is the immune complex;

[0099] 2. After coating the ELISA plate with C1q, incubate the serum of patients with rheumatoid arthritis, normal human serum, and peroxide reductase IV immune complexes prepared in vitro, recombinan...

Embodiment 3

[0103] Example 3 Application of High Performance Liquid Chromatography to Confirm the Existence of Peroxide Reductase IV Circulating Immune Complex

[0104] 1. Basic principles

[0105] Circulating immune complex is a combination of antigen and antibody, its molecular weight and volume are larger than free antigen and antibody, so circulating immune complex can be separated from serum by high performance liquid chromatography. Furthermore, the specific substances existing in the form of immune complexes were confirmed in the large molecular weight components separated from the serum.

[0106] 2. Test steps

[0107] For the separation of serum components, refer to the literature method "Xiaoyan Zhao, Nwora Lance Okeke, Orr Sharpe, Franak M Batliwalla, Annette T Lee, Peggy P Ho, Beren H Tomooka, Peter K Gregersen and William H Robinson. Circulating immune complexes contain citrullinated fibrinogen in rheumatoid Arthritis.Arthritis Research&Therapy2008, 10:R94", obtained compon...

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Abstract

The invention relates to the field of medical science and particularly verifies the existence of a circulating immune complex of peroxide reductase IV, which can be used as a diagnosis marker of rheumatoid arthritis. A capturing agent can be used as a diagnosis reagent of the rheumatoid arthritis. The invention further introduces a diagnosis kit for detecting the content of the circulating immune complex of the peroxide reductase IV of claims; a solid-phase carrier of the diagnosis kit is covered by the capturing agent of the circulating immune complex; the diagnosis kit further comprises a detection reagent which is combined with the capturing agent and can be quantitative. The invention verifies the existence of the circulating immune complex of the peroxide reductase IV for the first time; the invention discloses a content detection method of the circulating immune complex of the peroxide reductase IV, which can accurately detect the content of the circulating immune complex in blood serum of a patient and is good in repeatability; the diagnosis capability of the circulating immune complex of the peroxide reductase IV in the rheumatoid arthritis is higher than that of a RF (Rheumatoid Factor) and anti-CCP (anti-Cyclic Citrullinated Peptide); the circulating immune complex of the peroxide reductase IV can also be used as the diagnosis marker of other diseases and is high in distinction degree.

Description

technical field [0001] The invention relates to the field of medicine, in particular to an immune complex and its application. Background technique [0002] Antigen-antibody complex (antigen-antibody complex), according to the definition of the National Committee for Scientific and Technical Terms, refers to the multimolecular complex formed by the reversible combination of antibodies and antigens by means of secondary bonds such as electrostatic forces, hydrogen bonds, and van der Waals forces. . It can activate complement, participate in immune regulation, and mediate type III hypersensitivity. Immune complex (IC) is a kind of antigen-antibody complex. According to the definition of the National Committee for Scientific and Technical Terms, it refers to the complex formed by the combination of antibody and soluble antigen. When the amount of antibody is sufficient to cross-link the antigen, large immune complexes are formed, which are cleared by cells expressing Fc recep...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N33/573G01N33/577
Inventor 张大文李玲
Owner CHONGQING LIANBAI BOCHAO MEDICAL EQUIP
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