A Single-Chain Fv Antibody Against Platelet Membrane Glycoprotein IIb/IIIa Obtained Using Phage Display Technology

Objective The purpose of this study was to obtain the high-affinity single-chain Fv antibody against platelet membrane glycoprotein(GP) Ⅱb/Ⅲa from anti-activity platelet phage display library. Methods The heavy-chain and light-chain variable region gene(VH and VL)repertoire of immunoglobulin were amplified from the speen cells mRNA by RT-PCR and joined by a DNA linker encoding peptide as a single-chain Fv(ScFv) fragment with overlap extension. These fragments were cloned into the phagemid pHEN1 and the phage display library was constructed. The affinity selection and ELISA were adopted for identification of specific phage antibody to GPⅡb/Ⅲa. Results After through 4 rounds of panning, the high affinity ScFv gene was obtained, the sequence was comfined with mouse antibody. Conclusion Phage display library of repertoire single chain antibody of anti activity platelet was successfully constructed, and gene of single chain antibody which have binding ability to activity platelet was screened.