Expression of soluble VEGF receptor 2 and characterization of its binding by surface plasmon resonance

Xianming Huang, Claudia Gottstein, Rolf A. Brekken, Philip E. Thorpe

Research output: Contribution to journalArticle

27 Scopus citations

Abstract

Vascular endothelial growth factor (VEGF) is an endothelial cell specific mitogen that induces angiogenesis in several pathological conditions. To block angiogenesis, soluble VEGF receptor can be used. In this study, we describe a method for high yield expression of soluble VEGF receptor 2 (sFlk-1) in a baculovirus expression system (30 mg purified sFlk-1 per L of insect cell supernatant). We also determined the binding constants for both human and mouse VEGF to the recombinant receptor by surface plasmon resonance. In this cell-free assay, under the given experimental conditions, the on-rate k(a) was 0.5-2.2 x 106 M-1 s-1 and the off-rate k(d) was 2-4 x 10-4 s-1 (K(D) = 2-6 x 10-10 M). To our knowledge this is the first study to report on- and off-rates for the VEGF:sFlk-1 interaction. Heparin was not required for the binding of VEGF to sFlk-1 in this assay. The obtained values will serve as baseline parameters for the design of improved versions of recombinant soluble VEGF receptor.

Original languageEnglish (US)
Pages (from-to)643-648
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume252
Issue number3
DOIs
StatePublished - Nov 27 1998

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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