Chemiluminescence immunoassay for vascular endothelial growth factor (vascular permeability factor) in tumor-tissue homogenates.
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.
Pharmaceuticals Research Laboratories, Ciba-Geigy, Basle, Switzerland. jean-marc.schlaeppi@chbs.mhs.ciba.com
J M
JM
Schlaeppi
U
U
Eppenberger
G
G
Martiny-Baron
W
W
Küng
D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D001943:Breast Neoplasms / Q000737:chemistry*; D003600:Cytosol / Q000737:chemistry; D016228:Endothelial Growth Factors / Q000032:analysis* / Q000276:immunology; D005260:Female; D006801:Humans; D007118:Immunoassay / Q000379:methods* / Q000706:statistics & numerical data; D008163:Luminescent Measurements; D008222:Lymphokines / Q000032:analysis* / Q000276:immunology; D010051:Ovarian Neoplasms / Q000737:chemistry*; D011379:Prognosis; D011994:Recombinant Proteins / Q000276:immunology; D012680:Sensitivity and Specificity; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
D000911:Antibodies, Monoclonal; D016228:Endothelial Growth Factors; D008222:Lymphokines; D011994:Recombinant Proteins; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors