1. Pathogenicity Anti-Japanese 2-GPI antibody inhibits the anticoagulant activity of activated protein C, p2-GPI, antithrombin Ⅲ, inhibits factor Ⅻ-dependent fibrinolysis, increases adhesion molecule expression, and enhances endothelial cell procoagulant activity, while Mediates thrombosis. Oxidative modification of LDL is often considered to be a key factor in the formation of atherosclerosis, β2 GPI can specifically bind oxidized lipoproteins, such as oxidized VLDL, LDL, HDL, anti-p2-GPI can be complexed with β2-GPI-oxidized lipoprotein Combine. This immune response caused by anti-β2-GPI may be related to atherosclerosis.
2. Detection method: ELISA method: add 50μl 10μg / m1 per day 2-GPI (diluted with pH7.4 HEPES buffer), 4C binding 16-20h; the coated plate was washed three times with 200ul PB ~ Tween20, and then 200 ~ 1 HEPES buffer containing 3% gelatin was incubated at room temperature and washed after 1h; then, the sample was diluted with 50ra per well, placed at room temperature for 1h, washed plate; add 100tA HRP labeled mouse anti-human IgG or goat anti-mouse IgG, room temperature for 1h; after TMB developed for 10 minutes, stop solution was added. Read absorbance value at 450nm wavelength.
The solid phase antigens used in ELISA are:
①β2-GPI combined with cardiolipin coated polyethylene plate;
②Polyoxygenated polystyrene board adsorbed with β2-GPI. When using cardiolipin-coated plates, wells without β2-GPI should be set as a control; meanwhile, antibody detection results with and without β2-GPI are compared to evaluate the dependence of antibodies on p2-GPI.
3. Clinical Significance Anti-phospholipid antibody syndrome is a disease involving multiple systems that is characterized by a continuous increase in phospholipid antibodies, arteriovenous thrombosis, thrombocytopenia, and repeated spontaneous abortions. Anti-phospholipid antibodies mainly include anti-cardiolipin antibodies, anti-β2-glycoprotein I antibodies, anti-phosphatidylserine antibodies and lupus anticoagulants. Anti-p2-GPI antibodies seem to be the main autoantibodies in anti-phospholipid antibody-positive APS patients. Therefore, the detection of anti-p2-GPI antibodies in serum is particularly important for the diagnosis of APS.
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