In vitro functional characterization of human anti-bullous pemphigoid 180 (BP180) single-chain Fv antibody / 中华皮肤科杂志

ABSTRACT

Objective To characterize the function of human anti-BP180 single-chain Fv antibody (scFv) in vitro. Methods The IgG autoantibodies against BP180 were purified by affinity chromatography from the sera of patients with BP. The inhibitive effect of previously constructed anti-BP180 scFv on the binding of anti-BP180 IgG autoantibodies to the recombinant NC16A domain of human BP180 antigen was observed by competitive ELISA, competitive immunofluorescence assay and competitive inhibition test for complement activation. Results As ELISA revealed, the scFv significantly inhibited the binding of anti-BP180 IgG autoantibodies to the corresponding antigen (P ＜ 0.01 ), and the inhibitive effect was dose-dependent within the concentration range from 0 to 60 μg/ml. The inhibitive rate peaked at 69.50%. The deposition of anti-BP180-IgG autoantibodies in basement membrane zone and the IgG autoantibody-mediated complement C3 activation were both suppressed by the scFv of 40 μg/ml. Conclusion The genetically engineered anti-BP180 scFv has a certain inhibitive effect on the binding of BP-IgG autoantibodies to BP180 antigen and on the subsequent complement activation in vitro.