Identification and anti-tumor activities analysis of recombinant humanized anti-HGF monoclonal antibody

YAO Zhiwei; LI Dan; MA Baorui; ZHANG Jie; SUN Chunyun; XIE Liangzhi; GAI Wenlin; SHENG Wang

doi:10.13431/j.cnki.immunol.j.20170078

Abstract：This study was designed to identify humanized monoclonal antibodies against HGF and investigate their anti-tumor activities and pharmacokinetics characteristics in mice.The concentration,molecular weight and purity of the antibodies developed by genetic engineering technology were evaluated by UV and SDS-PAGE.The regulatory effect of the antibodies on U87MG cells proliferation was further tested by MTT;the binding abilities of the antibodies to HGF were measured by indirect ELISA;the inhibition of binding of HGF to its receptor by developed antibody was evaluated by ELISA.The regulatory effect of the antibodies on PC-3 cell migration was tested by transwell assay;the in vivo antitumor effects of the antibodies were evaluated by using established xenograft animal model of U87MG.Pharmacokinetics characteristics of the antibodies were evaluated by intravenous injection in mice.Date showed that the purity of two antibodies reached more than 90％,and both of them had strong ability to inhibit cell proliferation of U87MG.There was no significant difference in antibody activity as compared with non-humanization.Both antibodies had a strong binding ability to HGF and could prevent the binding of HGF to c-Met.Cell migration experiments showed that the two antibodies had the ability to inhibit the promoting effect of HGF on PC-3 cell migration.In U87MG xenograft model,antibody HE19-12 showed better anti-tumor activity than HD19-27.The half-lifes of HE19-12 and HD19-27 were 423.3 and 342.5 h,respectively.In conclusion,two humanized antibodies targeting HGF/C-Met pathway are prepared,which demonstrate good therapeutic potential against glioma with optimal pharmacokinetics characteristics.