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

D J Binnema, J J van Iersel, G Dooijewaard

    Research output: Contribution to journalArticleAcademicpeer-review

    Abstract

    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.

    Original languageEnglish
    Pages (from-to)569-577
    JournalThrombosis and Haemostasis
    Volume43
    Issue number5
    DOIs
    Publication statusPublished - 1 Sept 1986

    Keywords

    • Animals
    • Antibodies
    • Antigen-Antibody Complex
    • Aorta
    • Carcinoma, Squamous Cell
    • Cell Line
    • Cercopithecus aethiops
    • Culture Media
    • Endothelium
    • Enzyme-Linked Immunosorbent Assay
    • Fibroblasts
    • Humans
    • Isoflurophate
    • Kidney
    • Liver
    • Melanoma
    • Reference Values
    • Urokinase-Type Plasminogen Activator
    • Journal Article
    • Research Support, Non-U.S. Gov't

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