To prepare human antibody to rabies virus with high neutralizing potency using ribosome display technology. The immunoglobulin heavy and light chain variable (VH, VL) genes were prepared with the peripheral blood lymphocytes from three volunteers immunized with rabies virus vaccine by PCR. The genes encoding scFv fragments were prepared by randomly combining VH and VL genes by SOE PCR. Rabies virus glycoprotein (RVGp) specific scFv genes were selected over five cycles of ribosome display. The isolated scFv genes were cloned into pET22b(+)/BL21(DE3), from which soluble scFv fragments were prepared. The expressed products of selected clones were analyzed by ELISA for isolating the positive clones. To improve the stability of obtained scFvs, VH-Lc-VK were constructed and characterized. A human scFv gene library with 6.2×10¹² numbers used for ribosome display was constructed. Among the 180 selected clones, four clones RB24, RB71, RB109 and RB156 which exhibited the highest ELISA signals were isolated. The analysis of their sequences showed that they were new human immunoglobulin V genes to RVGp. And the reconstructed VH-Lc-VK antibody fragments can recognize RVGp specifically and antagonize the cytolytic effect of rabies virus on Vero cells. The prepared VH-Lc-VK fragments to RVGp will be useful for preparing engineering antibodies with high affinity against rabies virus. Ribosome display is a rapid means of generating fully human antibody fragments in vitro.