Construction and expression of mouse/human chimeric Fab of mAb 1A8 against HFRS virus on phage

AIM To construct and to express the mouse/human chimeric Fab antibodies on phage. METHODS Mouse/human chimeric heavy chain Fd (Fd fragment) and light chain genes were generated by ligation of V H gene of mouse mAb 1A8 against HFRS virus with human heavy chain CH1 gene and V L gene with human C κ gene, respectively. The chimeric Fab expression vector was constructed by cloning the chimeric heavy chain Fd and light chain genes into pComb3. Fab antibodies were expressed on phage by superinfection with helper phage VCSM13, and the binding activity of antibodies was detected by indirect sandwich ELISA. RESULTS Mouse/human chimeric Fab expression vector was constructed successfully, which was identified by restriction enzymes digestion. The supernatant of superinfection could be detected that having activity of binding HFRS virus antigen by the indirect sandwich ELISA. CONCLUSION Mouse/human chimeric Fab antibody expression vector was constructed and expressed on the surface of phage successfully. The Fab antibodies had binding activity against HFRS virus antigen.