Quantitation of urokinase antigen in plasma and culture media by use of an ELISA - PubMed (original) (raw)

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

D J Binnema et al. Thromb Res. 1986.

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.

PubMed Disclaimer

Cited by

Reprogramming urokinase into an antibody-recruiting anticancer agent.
Jakobsche CE, McEnaney PJ, Zhang AX, Spiegel DA. Jakobsche CE, et al. ACS Chem Biol. 2012 Feb 17;7(2):316-21. doi: 10.1021/cb200374e. Epub 2011 Nov 18. ACS Chem Biol. 2012. PMID: 22098560 Free PMC article.
Activated protein C inhibits local coagulation after intrapulmonary delivery of endotoxin in humans.
van der Poll T, Levi M, Nick JA, Abraham E. van der Poll T, et al. Am J Respir Crit Care Med. 2005 May 15;171(10):1125-8. doi: 10.1164/rccm.200411-1483OC. Epub 2005 Mar 4. Am J Respir Crit Care Med. 2005. PMID: 15750041 Free PMC article. Clinical Trial.
Local activation of coagulation and inhibition of fibrinolysis in the lung during ventilator associated pneumonia.
Schultz MJ, Millo J, Levi M, Hack CE, Weverling GJ, Garrard CS, van der Poll T. Schultz MJ, et al. Thorax. 2004 Feb;59(2):130-5. doi: 10.1136/thorax.2003.013888. Thorax. 2004. PMID: 14760153 Free PMC article.
Contribution of plasminogen activators and their inhibitors to the survival prognosis of patients with Dukes' stage B and C colorectal cancer.
Ganesh S, Sier CF, Heerding MM, van Krieken JH, Griffioen G, Welvaart K, van de Velde CJ, Verheijen JH, Lamers CB, Verspaget HW. Ganesh S, et al. Br J Cancer. 1997;75(12):1793-801. doi: 10.1038/bjc.1997.306. Br J Cancer. 1997. PMID: 9192984 Free PMC article.
Modified proenzymes as artificial substrates for proteolytic enzymes: colorimetric assay of bacterial collagenase and matrix metalloproteinase activity using modified pro-urokinase.
Verheijen JH, Nieuwenbroek NM, Beekman B, Hanemaaijer R, Verspaget HW, Ronday HK, Bakker AH. Verheijen JH, et al. Biochem J. 1997 May 1;323 ( Pt 3)(Pt 3):603-9. doi: 10.1042/bj3230603. Biochem J. 1997. PMID: 9169591 Free PMC article.

Quantitation of urokinase antigen in plasma and culture media by use of an ELISA - PubMed (original) (raw)