CHEMILUMINESCENCE IMMUNOASSAY FOR VASCULAR ENDOTHELIAL GROWTH-FACTOR (VASCULAR-PERMEABILITY FACTOR) IN TUMOR-TISSUE HOMOGENATES

We developed a two-site chemiluminescence immunoassay for human vascul ar endothelial growth factor (VEGF). The assay recognized both VEGF(12 1) and VEGF(165) isoforms, but had no detectable cross-reactivity with platelet-derived growth factor or placenta growth factor. The range o f detection was between 30 ng/L and 30 mu g/L VEGF. Inter- and intraas say variations were 8.2-8.3% and 7.2-7.6%, respectively. VEGF concentr ations were measured in the cytosolic extracts of 45 ovarian and 142 p rimary 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 to tal protein, range 0-12.3 ng/mg). In 32 and 7 extracts of normal breas t tissues adjacent and distant to the tumors, respectively, VEGF conce ntrations were significantly much lower (P <0.0001). The detection of substantial amounts of VEGF in two invasive tumors (compared with norm al tissues) suggests that the assay should be a useful tool for invest igating the prognostic value of VEGF in breast and ovarian carcinomas and for selecting patients for future anti-VEGF therapy.