Objective:To construct human single chain variable fragment (scFv) antibody library using phage display technology,and screen out special antibodies against the glycoprotein of rabies virus(RABVG) . Methods:The total RNA was isolated from the donors vaccinated with rabies vaccines and was used to amplify VH and VL genes by RT-PCR. VH and VL genes were joined together by overlap PCR to form scFv. The purified scFv gene repertoires were cloned into the phage vector pComb3XSS to construct the primary phage library. Panning against RABVG was performed for 5 rounds and the phage library was identified. The positive recombinant phages identified by ELISA were used to infect E.coli TOP10F′ for soluble scFv. Results:A recombinant phage antibody library against RABV was successfully constructed. 43 clones were found to bind to RABVG. 15 clones that had higher A450 values were detected with DNA sequencing and 3 of them were confirmed. The scFv gene was transformed into E.coli TOP10F′ for expression and purification. The purified scFv was proved to specifically bind to RABVG from the ELISA results. Conclusion:A human phage-display antibody library has been successfully constructed, and the selected scFv fragment can specifically bind to RABVG. It could be used in further studies of generation of human anti-RABVG antibodies for clinical application.