title: Phage-derived fully human antibody scFv fragment directed against human vascular endothelial growth factor receptor 2 blocked its interaction with VEGF.
abstract: Vascular endothelial growth factor receptor 2 (VEGFR-2) plays a critical role in tumor angiogenesis. None therapeutic antibodies targeting VEGFR-2 are available in clinical use. Herein, we describe the screening of a new single-chain antibody fragment (scFv) targeting extracellular domain 3 of human VEGFR-2 (kinase insert domain-containing receptor [KDR]3) from Griffin phage display scFv library. A comprehensive sequence analysis was performed to assign the framework and complementary-determining regions. The scFv exerted particular binding sites to KDR3 on molecular docking, and the binding affinity was further convinced by binding analysis both in quantitative ELISA and real-time kinetic determination by biosensors (K(D) = 40 nM). Finally, the scFv was revealed to inhibit VEGF-stimulated proliferation of human umbilical vein endothelial cells (HUVECs; IC(50) = 5 nM) and to inhibit HUVEC migration significantly at 17 nM. Taken together, our results indicate that we have successfully isolated a scFv which differentially recognizes KDR3 and has potential clinical applications in the treatment of angiogenesis related diseases.
authors.affiliation: Dept. of Molecular Biology, State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 210009, China. juancpu@126.com
authors.forename: Juan
authors.initials: J
authors.lastname: Zhang
authors.forename: Haixin
authors.initials: H
authors.lastname: Li
authors.forename: Xiuyun
authors.initials: X
authors.lastname: Wang
authors.forename: Haidi
authors.initials: H
authors.lastname: Qi
authors.forename: Xiaoniu
authors.initials: X
authors.lastname: Miao
authors.forename: Tao
authors.initials: T
authors.lastname: Zhang
authors.forename: Guosheng
authors.initials: G
authors.lastname: Chen
authors.forename: Min
authors.initials: M
authors.lastname: Wang
mesh_terms: D000595:Amino Acid Sequence; D001435:Bacteriophages / Q000235:genetics / Q000378:metabolism; D002465:Cell Movement; D049109:Cell Proliferation; D015536:Down-Regulation; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D019151:Peptide Library; D017434:Protein Structure, Tertiary; D057127:Single-Chain Antibodies / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D042461:Vascular Endothelial Growth Factor A / Q000276:immunology*; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000737:chemistry / Q000276:immunology*
chemical_list: D019151:Peptide Library; D057127:Single-Chain Antibodies; D042461:Vascular Endothelial Growth Factor A; D040301:Vascular Endothelial Growth Factor Receptor-2