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Detection method of human myelin oligodendrocyte glycoprotein autoimmune antibody

A technology of autoimmune antibodies and detection methods, which is applied to the detection field of human myelin oligodendrocyte glycoprotein autoimmune antibodies, can solve the problems of serious specificity and sensitivity, inability to specifically bind antibodies, etc. problems, to achieve important research and practical value, good specificity, and high detection efficiency.

Active Publication Date: 2019-04-23
广州敏特生物技术有限公司
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Problems solved by technology

[0003] Early detection methods for MOG autoimmune antibodies mainly used western blot and enzyme-linked immunosorbent assay, but the latest research at home and abroad shows that MOG autoantibodies from clinical patients need to specifically bind to MOG antigens with spatial conformation and glycosylation modification , and the MOG antigen prepared by the method adopted in the early stage does not have the correct spatial conformation or glycosylation modification, so it cannot specifically bind to the patient-derived MOG autoantibody, and there are serious problems in specificity and sensitivity

Method used

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  • Detection method of human myelin oligodendrocyte glycoprotein autoimmune antibody
  • Detection method of human myelin oligodendrocyte glycoprotein autoimmune antibody
  • Detection method of human myelin oligodendrocyte glycoprotein autoimmune antibody

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Experimental program
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Embodiment

[0019] 1 Construction of expression vector

[0020] Try a variety of MOG expression vectors, including pcDNA3.1-MOG, pcDNA3.1-MOG-GFP (expressing MOG and GFP fusion protein), pEGFP-IRES-MOG, etc., by screening and comparing the level of MOG antigen expressed on the cell surface, the expression vector pEGFP-IRES-MOG(expression vector2)( figure 1 ) MOG antigen expressed on the cell surface was significantly higher than other expression vectors.

[0021] The pEGFP-IRES-MOG vector was constructed by inserting the internal ribosome entry site sequence (IRES) downstream of the green fluorescent protein (GFP) gene in the eukaryotic expression vector pEGFP vector (with CMV promoter), and cloning after the IRES The cDNA sequence encoding the full-length human MOG was obtained. The human calnexin gene was cloned into the pcDNA3.1 vector to obtain the pcDNA-clanexin expression vector.

[0022] 2 chaperone calnexin promotes membrane expression of MOG antigen

[0023] MOG autoimmune an...

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Abstract

The invention discloses a detection method of a self immune antibody. The method comprises the following steps of performing cotransfection cells by expression vectors of allergen protein of an antibody to be detected and a chaperone protein expression vector; after the transfected cells are fixed and after washing, adding serum to be tested for incubation; adding second antibodies for incubation; then, performing analysis, wherein the antigen protein of the antibody to be detected is MOG protein. The detection method has the characteristics of high detection efficiency, high specificity and high sensitivity. The common problem of false positive or false negative detection result in the immunoblotting and enzyme linked immunosorbent assay are successfully solved; very important study and practical values are realized.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a detection method of human myelin oligodendrocyte glycoprotein autoimmune antibody. Background technique [0002] Myelin oligodendrocyte glycoprotein (MOG) is a myelin protein component that is specifically produced by oligodendrocytes and exists in the outermost layer of myelin in the central nervous system (CNS). The physiological functions of MOG include the constituent proteins of myelin, cell surface adhesion molecules mediating the interaction between myelin and extracellular matrix, regulating the stability of microtubules and participating in the activation of the complement system, etc. Studies at home and abroad have shown that autoimmune antibodies against MOG can be detected in a variety of central nervous system inflammatory demyelinating diseases, such as multiple sclerosis (MS), acute myelitis (AM) and neuromyelitis optica (NMO). . And MOG antibody-positive p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68
CPCG01N33/6893G01N2800/285
Inventor 彭立胜邱伟王施思陆正齐
Owner 广州敏特生物技术有限公司
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