Production of an anti-idiotypic antibody single chain variable fragment vaccine against Edwardsiella tarda

• Main Authors: Hong Qin, Xiaohang Jin, Weiquan Huang, Yulin Liu
• Other Authors: The Pennsylvania State University CiteSeerX Archives
• Format: Text
• Language: English
• Subjects: Received; Red drum; Sciaenops ocellatus
• Online Access: http://citeseerx.ist.psu.edu/viewdoc/summary?doi=10.1.1.621.1747; http://www.abbs.info/pdf/42-02/06.pdf

Edwardsiella tarda is the pathogen responsible for edward-siellosis, a serious infectious disease of freshwater and marine fish species, and currently recognized to be the species pathogenic for human. An anti-idiotypic mono-clonal antibody (mAb), 1E11, has been developed. It mimics the protective epitope of E. tarda and can prevent fish from infection of E. tarda. In this study, the correct variable heavy (VH) and variable light (VL) genes were obtained from 1E11 by using bioinformatics methods, and a 15 amino acid (Gly4Ser)3 linker was used to hold the two V domains together for the construction of VL– linker–VH form of single chain variable fragment (scFv) gene. Then, the scFv was subcloned into the vector pET-28a, expressed in the Escherichia coli BL21 cells, and identified by SDS–PAGE and western blotting. Red drum (Sciaenops ocellatus L.) weighing about 50 g was sub-jected to challenge with different E. tarda strains after 4 weeks followed by vaccination, the mortality rates and relative percentage survival were recorded and calculated, and the survival rate of fish in the scFv subgroups was obviously higher than that of control subgroups (P < 0.01). Enzyme-linked immunosorbent assay results show that after 4 weeks of post-vaccination, the level of specific antibody in fish sera of scFv groups was significantly higher than control groups. This study indicates that the recombinant antibody scFv was successfully developed, and it may serve as an effective vaccine candidate against E. tarda.