Abstract
A single-chain antibody phage display library was constructed from spleen cells of mice immunized with a soluble form of a human vascular endothelial growth factor (VEGF) receptor, kinase insert domain-containing receptor (KDR). After two rounds of biopanning, > 90% of the clones recovered were specifically reactive to KDR. Subsequent selection identified two clones that blocked VEGF binding to KDR. The clones were expressed in Escherichia coli and purified as soluble single-chain Fv (scFv) antibodies. The affinities of the scFv for binding to KDR were determined by BIAcore analysis (2.1 × 10-9–5.9 × 10-9 m). One scFv, p1C11, was shown to inhibit VEGF-induced KDR phosphorylation and VEGF-stimulated DNA synthesis in human umbilical vein endothelial cells. There is much experimental evidence to suggest that the VEGF/KDR/Flk-1 pathway plays an important role in tumor angiogenesis, a process that is essential for tumor growth and metastasis. The antibodies discussed here, which block VEGF binding to KDR, have potential clinical application in the treatment of cancer and other diseases where pathological angiogenesis is involved.
Footnotes
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↵1 To whom requests for reprints should be addressed, at Department of Molecular and Cell Biology, ImClone Systems, Inc., 180 Varick Street, New York, NY 10014. Phone: (212) 645-1405; Fax: (212) 645-2054; E-mail: zhenping@imclone.com.
- Received March 4, 1998.
- Accepted June 16, 1998.
- ©1998 American Association for Cancer Research.
Volume 58, Issue 15
