Construction Of Naive Channel Catfish ScFv Phage Display Antibody Library And Preliminary Identificiation

Antibodies play an important role in biological research, the discovery of phage antibody library technology makes the preparation of antibodies more convenient. It is still blank for Antibody library technology applied to the research of aquatic biology, this paper constructed natural channel catfish against single-stranded Phage library were studied.Recombinant Phage Anibody System,the genes of VH and VL were extracted from the spleen cells of non-immunized hannel catfish lymphocytes cells o then the VH and VL were joined together with a linker DNA by splicing overlap extension PCR,and a desired fragment (ScFv) was generated.After the ScFv genes were digested with Sfil and Notl,the arget fragment were ligated into the phagemid vector pCANTAB-5E and transformed into competent Ecoli TGI cells.The recombinants were grown on the amp plate.Plasmid electrophoresis showed that there were insert fragments and the target fragment (1300bp) were amplified by PCR.The result showed that the natural channel catfish against single-stranded Phage library were successfully constructed and it contain 5.8×105 different clones.OVA-SMZ was used for panning and screening ScFv antibody from the constructed natural channel catfish against single-stranded Phage library.Clones with good reactivity with pig OVA-SMZ in ELISA selected soluble ScFv antibodies were produced and the expression conditions were optimized.Furthermore,the characteristics of OVA-SMZ ScFvs were determined by ELISA.The products were purified by saturated animonlum sulfite and ion-exchange chromatography.Our study plays an important role for the antibody preparation and the exploration of new biological products. which opens a new filed for disease treatment and detection of drug residues on aquatic oranisms.