A fluorescence polarization assay for identifying ligands that bind to vascular endothelial growth factor.

Publications // Sheibani Lab // Jul 01 2008

PubMed ID: 18413228

Author(s): Peterson KJ, Sadowsky JD, Scheef EA, Pal S, Kourentzi KD, Willson RC, Bresnick EH, Sheibani N, Gellman SH. A fluorescence polarization assay for identifying ligands that bind to vascular endothelial growth factor. Anal Biochem. 2008 Jul 1;378(1):8-14. doi: 10.1016/j.ab.2008.03.043. Epub 2008 Mar 28. PMID 18413228

Journal: Analytical Biochemistry, Volume 378, Issue 1, Jul 2008

Vascular endothelial growth factor (VEGF) is a homodimeric proangiogenic protein that induces endothelial cell migration and proliferation primarily through interactions with its major receptors, VEGFR-1 and VEGFR-2. Inhibitors of one or both of these VEGF-receptor interactions could be beneficial as therapeutics for diseases caused by dysfunctional angiogenesis (e.g., cancer). Others have reported small peptides that bind to the VEGF dimer at surface regions that are recognized by the receptors. Here we report the development of a fluorescence polarization assay based on the binding to VEGF of a derivative of one of these peptides that has been labeled with BODIPY-tetramethylrhodamine (BODIPY(TMR)). This 384-well format assay is tolerant to dimethyl sulfoxide (DMSO, up to 4% [v/v]) and has a Z’ factor of 0.76, making it useful for identifying molecules that associate with the receptor-binding surface of the VEGF dimer.