J. Z. and H. L. contributed equally to this article
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Bioseparations and Downstream Processing
Phage-derived fully human antibody scFv fragment directed against human vascular endothelial growth factor receptor 2 blocked its interaction with VEGF
Article first published online: 18 JUN 2012
DOI: 10.1002/btpr.1559
Copyright © 2012 American Institute of Chemical Engineers (AIChE)
Additional Information
How to Cite
Zhang, J., Li, H., Wang, X., Qi, H., Miao, X., Zhang, T., Chen, G. and Wang, M. (2012), Phage-derived fully human antibody scFv fragment directed against human vascular endothelial growth factor receptor 2 blocked its interaction with VEGF. Biotechnol Progress, 28: 981–989. doi: 10.1002/btpr.1559
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J. Z. and H. L. contributed equally to this article
Publication History
- Issue published online: 7 AUG 2012
- Article first published online: 18 JUN 2012
- Accepted manuscript online: 11 MAY 2012 10:09AM EST
- Manuscript Revised: 12 APR 2012
- Manuscript Received: 19 DEC 2011
Funded by
- Natural Science Fund Committee project. Grant Numbers: NSFC81072561, NSFC81102364
- Project Program of State Key Laboratory of Natural Medicines(China Pharmaceutical University). Grant Number: JKGP201101
- Colleges and Universities in Jiangsu Province Plans to Graduate Research and Innovation Projects. Grant Number: 2011 CXZZ11_0818
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Keywords:
- vascular endothelial growth factor receptor 2 (VEGFR-2);
- single-chain antibody fragment (scFv);
- phage display;
- angiogenesis
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 (KD = 40 nM). Finally, the scFv was revealed to inhibit VEGF-stimulated proliferation of human umbilical vein endothelial cells (HUVECs; IC50 = 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. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 28: 981–989, 2012