抗鹅细小病毒噬菌体单链抗体文库的构建研究

o construct a library of single chain variable fragment (scFv) antibody against goose parvovirus （GPV）using phage display and getting the scFv antibody against GPV with high speciality and affinity by bio-panning. Total RNA was extracted from the spleen cell of the immuned goose，followed RT-PCR using random primer. The heavy-chain and light-chain variable region genes (VH and VL) repertoire of immunoglobulin were amplified respectively by PCR. Then they were linked by a DNA linker encoding (G1y4Ser)3as VH-linker-VL sequence forming scFv by PCR. These fragments were cloned into the phagemid pCANTAB5E and the recombinant phagemids were transformed to susceptible Escherichia coli TGl by electroporation. After rescuing with helper phage M13KO7, a library of phage scFv antibody was got. GPV was coated on 96 well plate to perform affinity enrichment panning, and to test the bio-activity of the scFv. IgBlast results revealed that the Vλ gene sharing 91.5 % identity with those of human immunoglobulin germline genes, while the VH gene shared only 75% identity.The recombination rate of the recombinant plasmids scFv-pCANTAB5E is 90%, the titer of the phage antibody library was about 1.2 106pfu. The goose special scFv phage library against GPV was constructed successfully, and it establishes the foundation for applications of the phage display technique and studing on location diagnose in vivo.