Chemiluminescence immunoassay for vascular endothelial growth factor (vascular permeability factor) in tumor-tissue...

We developed a two-site chemiluminescence immunoassay for human vascular endothelial growth factor (VEGF). The assay recognized both VEGF121 and VEGF165 isoforms, but had no detectable cross-reactivity with platelet-derived growth factor or placenta growth factor. The range of detection was between 30 ng/L and 30 micrograms/L VEGF. Inter- and intraassay variations were 8.2-8.3% and 7.2-7.6%, respectively. VEGF concentrations were measured in the cytosolic extracts of 45 ovarian and 142 primary breast tumors. The amount of VEGF in the ovarian tumors (median = 0.46 ng/mg total protein, range 0-15.8 ng/mg) was significantly (P = 0.03) higher compared with the breast tumors (median = 0.24 ng/mg total protein, range 0-12.3 ng/mg). In 32 and 7 extracts of normal breast tissues adjacent and distant to the tumors, respectively, VEGF concentrations were significantly much lower (P < 0.0001). The detection of substantial amounts of VEGF in two invasive tumors (compared with normal tissues) suggests that the assay should be a useful tool for investigating the prognostic value of VEGF in breast and ovarian carcinomas and for selecting patients for future anti-VEGF therapy.