Construction And Screen Of A Phage Display Library Of Repertoire Single Chain Antibody From Mouse Immunized With Crude Extracts Of Cysticercus Cellulosae

Hai Gang(Preventive Veterinary Medicine) Supervised by Professor Cai Xue-Peng and Yue ChengObjective Anti-Cysticercus cellulosae phage display library was constructed from mouse immunized with crude extract antigen of Cysticercus cellulosae ,and the high affinity ScFv to the antigen would be selected from the phage display library.Methods The heavy-chain and light-chain variable region (VH and VL) repertoire of immunogloblin 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 extention.These fragments were cloned into the phagemid pHENl and the phage display library was constructed. The affinity selection and ELISA were adopted for identification of specific phage antibody to crude extracts antigen of Cyticercus cellulosae. Results After 3 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-Cyticercus cellulosae was successfully constructed and gene of single chain antibody which have binding ability to crude extracts antigen of Cyticercus cellulosae was screened.