title: Development of humanized rabbit monoclonal antibodies against vascular endothelial growth factor receptor 2 with potential antitumor effects.
abstract: Vascular endothelial growth factor-A (VEGF-A) plays a critical role in physiologic and pathologic angiogenesis through its receptors especially through VEGFR2. The lack of cross-reactivity of monoclonal antibodies with human VEGFR2/mouse Flk-1 is a major obstacle in preclinical developments. In this study, using a unique hybridoma technique, we generated a panel of 30 neutralization anti-VEGFR2 rabbit monoclonal antibodies (RabMAbs) either blocking VEGF/VEGFR2 interaction or inhibiting VEGF-stimulated VEGFR2 tyrosine kinase phosphorylation. Among 18 RabMAbs with human/mouse VEGFR2 cross-reactivity, we humanized one lead candidate RabMAb by Mutational Lineage Guided (MLG) method and further demonstrated its potent inhibition of tumor growth in xenograft mouse model. Our study suggests that RabMAbs are highly relevant for therapeutic applications.
authors.affiliation: Department of Urology, Sir Run-Run Shaw Hospital, College of Medicine, Zhejiang University, Hangzhou, China. yanlanyu@gmail.com
authors.forename: Yanlan
authors.initials: Y
authors.lastname: Yu
authors.forename: Pierre
authors.initials: P
authors.lastname: Lee
authors.forename: Yaohuang
authors.initials: Y
authors.lastname: Ke
authors.forename: Yongke
authors.initials: Y
authors.lastname: Zhang
authors.forename: Jungang
authors.initials: J
authors.lastname: Chen
authors.forename: Jihong
authors.initials: J
authors.lastname: Dai
authors.forename: Mingzhen
authors.initials: M
authors.lastname: Li
authors.forename: Weimin
authors.initials: W
authors.lastname: Zhu
authors.forename: Guo-Liang
authors.initials: GL
authors.lastname: Yu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D061067:Antibodies, Monoclonal, Humanized / Q000008:administration & dosage / Q000494:pharmacology*; D057134:Antibodies, Neutralizing / Q000008:administration & dosage / Q000494:pharmacology; D000915:Antibody Affinity; D000918:Antibody Specificity; D000970:Antineoplastic Agents / Q000008:administration & dosage / Q000494:pharmacology*; D003429:Cross Reactions; D005260:Female; D057809:HEK293 Cells; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D020128:Inhibitory Concentration 50; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D008969:Molecular Sequence Data; D009389:Neovascularization, Pathologic / Q000276:immunology / Q000628:therapy; D010766:Phosphorylation; D011485:Protein Binding; D025941:Protein Interaction Mapping; D011817:Rabbits; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000037:antagonists & inhibitors* / Q000276:immunology; D023041:Xenograft Model Antitumor Assays
chemical_list: D061067:Antibodies, Monoclonal, Humanized; D057134:Antibodies, Neutralizing; D000970:Antineoplastic Agents; D042461:Vascular Endothelial Growth Factor A; D040301:Vascular Endothelial Growth Factor Receptor-2
keywords: Angiogenesis; Anti-angiogenic therapy; Rabbit monoclonal antibody; Vascular endothelial growth factor receptor 2