title: Antagonists to human and mouse vascular endothelial growth factor receptor 2 generated by directed protein evolution in vitro.
abstract: Using directed in vitro protein evolution, we generated proteins that bound and antagonized the function of vascular endothelial growth factor receptor 2 (VEGFR2). Binders to human VEGFR2 (KDR) with 10-200 nM affinities were selected by using mRNA display from a library (10(13) variants) based on the tenth human fibronectin type III domain (10Fn3) scaffold. Subsequently, a single KDR binding clone (K(d) = 11 nM) was subjected to affinity maturation. This yielded improved KDR binding molecules with affinities ranging from 0.06 to 2 nM. Molecules with dual binding specificities (human/mouse) were also isolated by using both KDR and Flk-1 (mouse VEGFR2) as targets in selection. Proteins encoded by the selected clones bound VEGFR2-expressing cells and inhibited their VEGF-dependent proliferation. Our results demonstrate the potential of these inhibitors in the development of anti-angiogenesis therapeutics.
authors.affiliation: Phylos, Inc., succeeded by Compound Therapeutics, Inc., 100 Beaver Street, Waltham, Massachusetts 02453, USA.
authors.forename: Elena V
authors.initials: EV
authors.lastname: Getmanova
authors.forename: Yan
authors.initials: Y
authors.lastname: Chen
authors.forename: Laird
authors.initials: L
authors.lastname: Bloom
authors.forename: Jochem
authors.initials: J
authors.lastname: Gokemeijer
authors.forename: Steven
authors.initials: S
authors.lastname: Shamah
authors.forename: Veena
authors.initials: V
authors.lastname: Warikoo
authors.forename: Jack
authors.initials: J
authors.lastname: Wang
authors.forename: Vincent
authors.initials: V
authors.lastname: Ling
authors.forename: Lin
authors.initials: L
authors.lastname: Sun
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D019020:Directed Molecular Evolution; D006801:Humans; D051379:Mice; D016296:Mutagenesis; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000037:antagonists & inhibitors* / Q000378:metabolism
chemical_list: D009842:Oligopeptides; D011994:Recombinant Proteins; D040301:Vascular Endothelial Growth Factor Receptor-2