The application discloses a kind of targeting cat calicivirus VP1-CDE
protein chicken source single-chain
antibody and its preparation and application, belong to
biotechnology field.The single-chain
antibody is selected from X2B2, X2E1 Or X2G5, its
heavy chain variable region and light chain variable region
amino acid sequence are respectively as shown in SEQ ID NO:1-6.The application is
prokaryotic expression and purified VP1-CDE
protein as
antigen immunization chicken, amplifies VH And VL
Gene and is spliced into scFv
gene by (Gly4Ser) 3
linker, constructs
library capacity not less than 5.25×10 7 CFU / mL
phage display library, specific single-chain
antibody clone is obtained by
solid-phase panning, and its binding activity is verified by
Western blot, ELISA,
indirect immunofluorescence test after eukaryotic expression and purification.The results show that the three single-chain antibodies can specifically bind VP1-CDE
protein and FCV
virus particles, and there is no cross reaction, which provides key experimental materials for
basic research of cat calicivirus, can be used for developing FCV rapid
diagnostic reagent or kit, also provides candidate molecules for the development of FCV infection treatment drugs, and has important theoretical significance and application value.