title: A sensitive and robust assay for urokinase and tissue-type plasminogen activators (uPA and tPA) and their inhibitor type I (PAI-1) in breast tumor cytosols.
abstract: uPA and PAI-1 are becoming established as amongst the most effective markers of poor prognosis for patients with node-negative breast cancer; tPA is an index of longer survival. This paper describes a sensitive ELISA for the measurement of uPA, tPA and PAI-1 in breast cancer cytosols. The structure of the assay involves coating Ab (sheep alpha-Chicken IgY), catching Ab (chicken alpha-analyte), tagging Ab (rabbit alpha-analyte) and detecting Ab (goat alpha-rabbit IgG) labelled with HRP. The assay has a high degree of accuracy and specificity. Comparison with the American Diagnostic kits shows the results' equivalence for PAI-1 and tPA. For uPA the results of the assay were twice as high. The assay is sensitive and relatively inexpensive. It is the first published assay to yield strictly comparative values for uPA, tPA and PAI-1 in tissue extracts and is readily subject to external quality control.
authors.affiliation: Department of Experimental & Chemical Endocrinology, Academic Hospital St Radboud, University of Nijmegen, The Netherlands.
authors.forename: N
authors.initials: N
authors.lastname: Grebenschikov
authors.forename: A
authors.initials: A
authors.lastname: Geurts-Moespot
authors.forename: H
authors.initials: H
authors.lastname: De Witte
authors.forename: J
authors.initials: J
authors.lastname: Heuvel
authors.forename: R
authors.initials: R
authors.lastname: Leake
authors.forename: F
authors.initials: F
authors.lastname: Sweep
authors.forename: T
authors.initials: T
authors.lastname: Benraad
mesh_terms: D000818:Animals; D014408:Biomarkers, Tumor / Q000032:analysis* / Q000276:immunology; D001706:Biopsy; D001943:Breast Neoplasms / Q000201:enzymology* / Q000473:pathology; D002645:Chickens; D003600:Cytosol / Q000201:enzymology / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006041:Goats; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D017395:Plasminogen Activator Inhibitor 1 / Q000032:analysis* / Q000276:immunology; D011817:Rabbits; D011933:Reagent Kits, Diagnostic; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D015842:Serine Proteinase Inhibitors / Q000032:analysis* / Q000276:immunology; D012756:Sheep; D010959:Tissue Plasminogen Activator / Q000032:analysis* / Q000276:immunology; D014568:Urokinase-Type Plasminogen Activator / Q000032:analysis* / Q000276:immunology
chemical_list: D014408:Biomarkers, Tumor; D007106:Immune Sera; D017395:Plasminogen Activator Inhibitor 1; D011933:Reagent Kits, Diagnostic; D015842:Serine Proteinase Inhibitors; D010959:Tissue Plasminogen Activator; D014568:Urokinase-Type Plasminogen Activator