Objective：To develop monoclonal antibodies（mAb）against human neuropilin 1（HuNRP1）and provide useful materials to inhibit tumor angiogenesis by targeting HuNRP1. Methods：Using lymphocyte hybridoma technique，8-week-old BALB/c mice were immunized with recombinant protein，b domain of human neuropilin 1（TF-HuNRP1b）derived from E. Coli，then splenocytes of the immunized mice were prepared and collected，with the addition of sp2/0 myeloma cells，all the mixed cells were fused. Positive hybridoma cells were screened by indirect immunofluorescent assay（IFA）established with native HuNRP1 expressed by breast tumor cell line MDA-MB-231. The specificity of mAb was identified by IFA，Western blot and flow cytometry（FCM）. The inhibitory ability of mAbs against tumor angiogenesis was identified by methyl thiazolyl tetrazolium（MTT）and chamber migration test. Results：One hybridoma cell line producing mAb against TF-HuNRP1b were obtained and named as 4F11. Western blot analysis showed that mAbs could recognize only recombinant HuNRP1b not the other proteins produced in BL21（DE3）（pCold-HuNRP1b）and BL21（DE3）（pCold）. The results of IFA and FCM showed that mAb could recognize native HuNRP1 expressed on human umbilical vein endothelial cells（HUVEC），tumor cell line MDA-MB-231 and HepG2. The results of MTT assay and chamber migration test further showed that mAb 4F11 had strong inhibitory effect on migration and proliferation of HUVEC. Conclusion：mAb against HuNRP1b was successfully obtained which will be useful in the functional study of HuNRP1 and provide good materials for the inhibition of tumor angiogenesis.