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

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

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.

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OrganisatieHanzehogeschool Groningen
LectoraatBiobased Ingredients and Materials
Gepubliceerd inThrombosis and haemostasis Schattauer, Vol. 43, Uitgave: 5, Pagina's: 569-577
Datum1986-09-01
TypeArtikel
ISSN0049-3848
DOI10.1016/0049-3848(86)90077-0
TaalEngels

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