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 Sep 1986

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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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