Development of a phage displayed disulfide-stabilized Fv fragment vaccine against Vibrio anguillarum.
Anti-idiotype monoclonal antibody 1E10 can mimic the protective epitope of Vibrio anguillarum and be used as vaccine to prevent fish infection of V. anguillarum. In this study, the variable heavy (V(H)) domain and variable light (V(L)) domain of mAb1E10 were cloned by RT-PCR and were linked to each other by a disulfide bond engineered at position 44 of V(H) and position 105 of V(L) that lie between structurally conserved framework positions. Mutated V(H) 44 and V(L) 105 were inserted into phagemid pCANTAB5E. When co-transfected by recombinant pCANTAB5E and helper phage M13KO7, the host Escherichia coli cells secreted disulfide-stabilized Fv fragment (dsFv) which displayed on the surface of filamentous phage. The binding specificity of the phage-displayed dsFv was proved by ELISA method. Protection experiment showed that Japanese flounders can develop high titer of antibody against the dsFv and survival ratio of vaccinated group was significantly different from control groups. Thus, this phage-displayed dsFv may be used as vaccine against V. anguillarum in fishery.
Xia
Yong-juan
YJ
Department of Histology and Embryology, Fourth Military Medical University, Xi'an 710032, China.
Wen
Wei-hong
WH
Huang
Wei-quan
WQ
Huang
Bao-cheng
BC
D000595:Amino Acid Sequence; D000818:Animals; D001428:Bacterial Vaccines / Q000737:chemistry / Q000276:immunology*; D003001:Cloning, Molecular; D004220:Disulfides / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D005393:Fish Diseases / Q000517:prevention & control*; D008222:Lymphokines / Q000737:chemistry / Q000276:immunology*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D019151:Peptide Library; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012795:Sialoglycoproteins / Q000737:chemistry / Q000276:immunology*; D016019:Survival Analysis; D014614:Vaccines, Synthetic / Q000276:immunology; D014733:Vibrio / Q000276:immunology*; D014735:Vibrio Infections / Q000517:prevention & control* / Q000662:veterinary*
D001428:Bacterial Vaccines; D004220:Disulfides; C089422:Fv protein, human; D008222:Lymphokines; D019151:Peptide Library; D012333:RNA, Messenger; D012795:Sialoglycoproteins; D014614:Vaccines, Synthetic