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dc.contributor.authorErdag, B.en_US
dc.contributor.authorKoray Balcioglu, B.en_US
dc.contributor.authorOzdemir Bahadir, A.en_US
dc.contributor.authorSerhatli, M.en_US
dc.contributor.authorKacar O.en_US
dc.contributor.authorBahar, A.en_US
dc.contributor.authorSeker, U.O.S.en_US
dc.contributor.authorAkgun, E.en_US
dc.contributor.authorOzkan, A.en_US
dc.contributor.authorKilic, T.en_US
dc.contributor.authorTamerler, C.en_US
dc.contributor.authorBaysal, K.en_US
dc.date.accessioned2016-02-08T09:50:15Z
dc.date.available2016-02-08T09:50:15Z
dc.date.issued2011en_US
dc.identifier.issn8854513en_US
dc.identifier.urihttp://hdl.handle.net/11693/21723
dc.description.abstractHuman vascular endothelial growth factor (VEGF) and its receptor (VEGFR-2/kinase domain receptor [KDR]) play a crucial role in angiogenesis, which makes the VEGFR-2 signaling pathway a major target for therapeutic applications. In this study, a single-chain antibody phage display library was constructed from spleen cells of mice immunized with recombinant human soluble extracellular VEGFR-2/KDR consisting of all seven extracellular domains (sKDR D1-7) to obtain antibodies that block VEGF binding to VEGFR-2. Two specific single-chain antibodies (KDR1.3 and KDR2.6) that recognized human VEGFR-2 were selected; diversity analysis of the clones was performed by BstNI fingerprinting and nucleotide sequencing. The single-chain variable fragments (scFvs) were expressed in soluble form and specificity of interactions between affinity purified scFvs and VEGFR-2 was confirmed by ELISA. Binding of the recombinant antibodies for VEGFR-2 receptors was investigated by surface plasmon resonance spectroscopy. In vitro cell culture assays showed that KDR1.3 and KDR2.6 scFvs significantly suppressed the mitogenic response of human umbilical vein endothelial cells to recombinant human VEGF 165 in a dose-dependent manner, and reduced VEGF-dependent cell proliferation by 60% and 40%, respectively. In vivo analysis of these recombinant antibodies in a rat cornea angiogenesis model revealed that both antibodies suppressed the development of new corneal vessels (p < 0.05). Overall, in vitro and in vivo results disclose strong interactions of KDR1.3 and KDR2.6 scFvs with VEGFR-2. These findings indicate that KDR1.3 and KDR2.6 scFvs are promising antiangiogenic therapeutic agents. © 2011 International Union of Biochemistry and Molecular Biology, Inc.en_US
dc.language.isoEnglishen_US
dc.source.titleBiotechnology and Applied Biochemistryen_US
dc.relation.isversionofhttp://dx.doi.org/10.1002/bab.61en_US
dc.subjectantiangiogenicen_US
dc.subjectcorneal angiogenesis assayen_US
dc.subjectHUVECen_US
dc.subjectphage displayen_US
dc.subjectsingle-chain variable fragment (scFv)en_US
dc.subjectvascular endothelial growth factor receptor-2 (VEGFR-2)en_US
dc.subjectAngiogenesisen_US
dc.subjectAntiangiogenicen_US
dc.subjectHUVECen_US
dc.subjectphage displayen_US
dc.subjectSingle chain variable fragmentsen_US
dc.subjectVascular endothelial growth factoren_US
dc.subjectAnimal cell cultureen_US
dc.subjectAssaysen_US
dc.subjectCell proliferationen_US
dc.subjectEndothelial cellsen_US
dc.subjectPeptidesen_US
dc.subjectPlants (botany)en_US
dc.subjectSurface plasmon resonanceen_US
dc.subjectAntibodiesen_US
dc.subjectvasculotropin antibodyen_US
dc.subjectvasculotropin receptor 2en_US
dc.subjectangiogenesisen_US
dc.subjectanimal cellen_US
dc.subjectanimal experimenten_US
dc.subjectarticleen_US
dc.subjectbinding affinityen_US
dc.subjectbiopanningen_US
dc.subjectcell proliferationen_US
dc.subjectcontrolled studyen_US
dc.subjectcorneaen_US
dc.subjectDNA fingerprintingen_US
dc.subjectDNA sequenceen_US
dc.subjectenzyme linked immunosorbent assayen_US
dc.subjecthumanen_US
dc.subjecthuman cellen_US
dc.subjecthuman tissueen_US
dc.subjectin vitro studyen_US
dc.subjectin vivo studyen_US
dc.subjectmaleen_US
dc.subjectmouseen_US
dc.subjectnonhumanen_US
dc.subjectphage displayen_US
dc.subjectprotein bindingen_US
dc.subjectprotein domainen_US
dc.subjectraten_US
dc.subjectspleen cellen_US
dc.subjectsurface plasmon resonanceen_US
dc.subjectAmino Acid Sequenceen_US
dc.subjectAngiogenesis Inhibitorsen_US
dc.subjectAnimalsen_US
dc.subjectBase Sequenceen_US
dc.subjectCell Proliferationen_US
dc.subjectCorneaen_US
dc.subjectDose-Response Relationship, Immunologicen_US
dc.subjectEnzyme-Linked Immunosorbent Assayen_US
dc.subjectHuman Umbilical Vein Endothelial Cellsen_US
dc.subjectHumansen_US
dc.subjectMaleen_US
dc.subjectMiceen_US
dc.subjectMice, Inbred BALB Cen_US
dc.subjectMolecular Sequence Dataen_US
dc.subjectPeptide Libraryen_US
dc.subjectRatsen_US
dc.subjectRecombinant Proteinsen_US
dc.subjectSingle-Chain Antibodiesen_US
dc.subjectSpleenen_US
dc.subjectSurface Plasmon Resonanceen_US
dc.subjectVascular Endothelial Growth Factor Aen_US
dc.subjectVascular Endothelial Growth Factor Receptor-2en_US
dc.titleIdentification of novel neutralizing single-chain antibodies against vascular endothelial growth factor receptor 2en_US
dc.typeArticleen_US
dc.departmentTUBITAK Marmara Research Center, Genetic Engineering and Biotechnology Institute, P.O. Box 21, 41470 Gebze, Kocaeli, Turkeyen_US
dc.departmentNational Nanotechnology Research Center, Institute of Material Science and Nanotechnology, Bilkent University, Turkeyen_US
dc.departmentMolecular Neurosurgery Laboratory, Institute of Neurological Sciences, Marmara University, Istanbul, Turkeyen_US
dc.departmentFaculty of Medicine, Department of Neurosurgery, Marmara University, Istanbul, Turkeyen_US
dc.departmentMaterials Science and Engineering Department, University of Washington, Seattle, WA, United Statesen_US
dc.departmentMedical Faculty, Department of Biochemistry, Dokuz Eylül University, Izmir, Turkeyen_US
dc.citation.spage412en_US
dc.citation.epage422en_US
dc.citation.volumeNumber58en_US
dc.citation.issueNumber6en_US
dc.identifier.doi10.1002/bab.61en_US


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