Objective:To generate human monoclonal antibodies (mAbs) against the rabies virus (RV) by using the mice carrying human immunoglobulin transloci for immunization and further to identify their characters. Methods:The human IgM transgenic mice were immunized with the inactivated RV(CTN strain) as immunogen. The human anti-RV mAbs were screened and produced by using the classical hybridoma technique in a combinatory high throughput screening (HTS) strategy. The fully human mAbs were identified via the double antibody sandwich enzyme-linked immunosorbent assay(ELISA),and the specificity of mAbs was determined by the indirect ELISA,Dot Blot and Western Blot. In addition,the affinity of mAbs was detected by BiaCore X-100 system. Results:Two anti-RV hybridoma cell lines 9E3 and 9F2,which can produce human IgM mAbs and recognize the CTN strain of RV specifically in the indirect ELISA and the Dot Blot assays,have been established. The results of Western Blot demonstrated that the two prepared mAbs specifically recognize glycoprotein(G protein) of the rabies virus. The equilibrium dissociation constants (KD) of 9E3 and 9F2 were 2.62 × 10-10 mol/L and 4.06 × 10-11 mol/L,respectively. Conclusion:Two anti-RV fully human IgM mAbs with specificity and high affinity were generated. The first-phase preparations of mAbs could be the bases for the screening immunoprophylaxis neutralized monoclonal antibody and immunotherapy on the rabies.