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 α-Chicken IgY), catching Ab (chicken α-analyte), tagging Ab (rabbit α-analyte) and detecting Ab (goat α-rabbit IgG) labelled with HRP. The assay has a high degree of accuracy and specificity. Comparison with the American Diagnostica 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.