Generation and characterization of recombinant neutralizing antibody Fab against rabies virus

摘要

Passive immune therapy and the using of rabies vaccine were considered to be an important part of blocking rabies virus. Some single chain antibodies (scFv) against rabies virus were selected by phage display techniques. The purpose of this study is to transform the scFv into Fab, and identify the biologic activity. The variable region genes, VH and VL, were amplified by PCR from anti-rabies virus scFv plasmids. The Fab gene was inserted into vector pcomb3XSS and transformed into E. coliTop10 for a great deal of soluble expression and purification. The Fab was purified by IMAC with FPLC system, and detected by western blot and indirect ELISA. Meanwhile, the fluorescent antibody virus neutralisation test (FAVN) showed Fab hasa high neutralizing activity against rabies virus strain CVS-11. This Fab fragment can be further modified to generate a full-length antibody for rabies therapy.