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

D J Binnema; J J van Iersel; G Dooijewaard

doi:https://doi.org/10.1016/0049-3848(86)90077-0

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

D J Binnema, J J van Iersel, G Dooijewaard

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Research output: Contribution to journal › Article › Academic › peer-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 language	English
Pages (from-to)	569-577
Journal	Thrombosis and haemostasis
Volume	43
Issue number	5
DOIs	https://doi.org/10.1016/0049-3848(86)90077-0
Publication status	Published - 1 Sep 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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@article{32c1b73ecbec405c86006db5d349ef02,

title = "Quantitation of urokinase antigen in plasma and culture media by use of an ELISA",

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.",

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",

author = "Binnema, {D J} and {van Iersel}, {J J} and G Dooijewaard",

year = "1986",

month = sep,

day = "1",

doi = "https://doi.org/10.1016/0049-3848(86)90077-0",

language = "English",

volume = "43",

pages = "569--577",

journal = "Thrombosis and haemostasis",

issn = "0340-6245",

publisher = "Schattauer",

number = "5",

}

TY - JOUR

T1 - Quantitation of urokinase antigen in plasma and culture media by use of an ELISA

AU - Binnema, D J

AU - van Iersel, J J

AU - Dooijewaard, G

PY - 1986/9/1

Y1 - 1986/9/1

N2 - 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.

AB - 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.

KW - Animals

KW - Antibodies

KW - Antigen-Antibody Complex

KW - Aorta

KW - Carcinoma, Squamous Cell

KW - Cell Line

KW - Cercopithecus aethiops

KW - Culture Media

KW - Endothelium

KW - Enzyme-Linked Immunosorbent Assay

KW - Fibroblasts

KW - Humans

KW - Isoflurophate

KW - Kidney

KW - Liver

KW - Melanoma

KW - Reference Values

KW - Urokinase-Type Plasminogen Activator

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - https://doi.org/10.1016/0049-3848(86)90077-0

DO - https://doi.org/10.1016/0049-3848(86)90077-0

M3 - Article

C2 - 3750278

VL - 43

SP - 569

EP - 577

JO - Thrombosis and haemostasis

JF - Thrombosis and haemostasis

SN - 0340-6245

IS - 5

ER -