Detection Of Anti-bFGF Autoantibodies In Autoimmune Patients

AIM: To investigate the existence of anti-bFGF autoantibodies in autoimmune patients. Collecting lymphocytes from the patients with anti-bFGF autoantibodies. This will be preparation for cloning the genes of the human immunoglobulin Fd chain and k chain, and selecting human anti-bFGF Fab.METHODS: The optimal conditions of pH value of carbonate buffer, the concentration of bFGF, blocking solution and the type of microtitre were established by ELISA. 359 blood samples of patients with autoimmune disease were analyzed with ELISA for IgG and IgM of anti-bFGF autoantibodies.RESUTS: The optimal conditions in indirect ELISA included pH 9.8 carbonate buffer as coating solution, 0.5 × 10^-3 μg/μL bFGF, Microtitter plates made in Jiangsu Huaxing glass & apparatus factory and the best blocking solutions contained gelatin(0.2%) and BSA(1%). These modifications reduced non-specific binding and improved assay sensitivity. The average values of anti-bFGF IgG in SLE, RA, DM were 0.194±0.059, 0.215 ±0.104, 0.239 ± 0.050, respectively. They were higher than the normal in 0.173 ± 0.02. The average values of anti-bFGF IgM in SLE, RA, DM, ITP were 0.206 ± 0.061, 0.234 ± 0.117, 0.248 ± 0.040, 0.225 ± 0.060, respectively. They were higher than the normal in 0.182 ± 0.037. High titer anti-bFGF autoantibodies could be found in some SLE, RA, kidney disease, DM, FOU and ITR The mean value of A₄₉₂ in anti-bFGF autoantibodies positive patients samples were higher than the doubled normal mean value with SD.CONCLUSION: The results revealed that the anti-bFGF autoantibody exist in autoimmune patients, which will be useful in research of the mechanism of autoimmune disease and human anti-bFGF antibody.