Objective:To prepare a human scFv-Fc fusion antibody against rabies virus and to analyze its binding activity. Methods:The human scFv-Fc prokaryotic expression vector was constructed. After optimizing prokaryotic expression system,the human scFv-Fc fusion antibody against rabies virus was expressed and purified. Purified humanized scFv-Fc antibody was confirmed its binding activity by binding to purified inactivated rabies virus. Results:After sequence analysis,the human scFv-Fc-pColdⅡprokaryotic expression vector was successfully established. Through transforming E.coli.BL21 (DE3),human scFv-Fc fusion antibody was expressed by optimized induction of IPTG at concentration of 0.5 mmol/L. The expression of scFv-Fc fusion antibody was increased and its molecular weight was 57 000. By purification,the human scFv-Fc fusion antibody was purified with excellent binding activity even in 512 times diluted concentration. Conclusion:We have successfully established a whole human anti-rabies virus scFv-Fc-pColdⅡ prokaryotic expression vector,and expressed and purified scFv-Fc fusion antibody. This fusion antibody can lay a foundation for developing novel antibody-targeted drugs of rabies prophylaxis after exposure.