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.
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[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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