Quantitation of urokinase antigen in plasma and culture media by use of an ELISA

D J Binnema, J J van Iersel, G Dooijewaard

    Onderzoeksoutput: ArticleAcademicpeer review

    Samenvatting

    An ELISA was set up using polyvinylchloride microtiter plates coated with rabbit anti-UK IgG's and affino-purified goat anti-UK IgG's as second antibody. Detection occurred with rabbit anti-goat IgG antibodies conjugated with alkaline phosphatase. The assay is specific for urokinase (UK) with a detection limit of 100 pg/ml sample. Tissue-type plasminogen activator, up to concentrations of 100 ng/ml, does not interfere. The assay measures the antigen of the inactive zymogen pro-UK, the active enzyme UK and the UK-inhibitor complex with equal efficiency and gives the total UK antigen present, irrespective of its molecular form. Culture media of fibroblasts, endothelial- and kidney cells showed, despite the absence of active UK, antigen levels of 1.2, 23 and 65 ng/ml, respectively. In human plasma the UK concentration was found to be 3.5 +/- 1.4 ng/ml (mean +/- SD, n = 54). The inter- and intra-assay variations were 20% and 6%, respectively.

    Originele taal-2English
    Pagina's (van-tot)569-577
    TijdschriftThrombosis and Haemostasis
    Volume43
    Nummer van het tijdschrift5
    DOI's
    StatusPublished - 1 sep. 1986

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