Construction and screening of a phage display library of repertoire single chain antibody from mouse immunized with P-selectin

Objective Anti-activity platelet phage display library was constructed from mouse immunized with P-selectin, which has been used in clinical usage. Methods The heavy-chain and light-chain variable region gene(VH and VL)repertoire of immunoglobulin were amplified from the spleen cell 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 P-selectin. Results After 5 rounds of panning, the high affinity ScFv gene was obtained, the sequence was confirmed with mouse antibody.Conclusion Phage display library of repertoire single chain antibody of anti P-selectin was successfully constructed, and gene of single chain antibody which have binding ability to P-selectin wass creened.