Objective:To construct a human scFv phage antibody library against rabies virus G protein,selected the single chain antibody against rabies virus G protein for different epitopes,and identificated its neutralizing activity. Methods:The human scFv phage antibody library against rabies virus G protein was constructed by using peripheral blood lymphocytes from 130 Rabies vaccine immune volunteers. Screening by the purified rabies virus G protein,the positive clones were identified and expressed,identified neutralizing activity by AMMS. Results:Through the prokaryotic expression system and purification of rabies virus G protein,the human anti rabies virus G protein scFv phage antibody library was constructed with about 5.0×107 size. The nucleic acid sequence analysis confirmed the insertion of a fragment is scFv. After 5 rounds of screening,we randomly picked 140 clones by phage-ELISA,and obtained 4 nucleic acid sequences in different scFv antibodies,purified and identified activity. We finally obtained 1 scFv antibody with neutralizing activity. Conclusion:We have successfully constructed a whole human anti-rabies virus G protein scFv antibody library,and obtained a whole human anti-rabies virus G protein scFv antibody with neutralizing activity from the library. It is established foundation of the new type of rabies virus vaccine for human immunization.