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Enzyme-linked immunosorbent assays for plasminogen activators

Authors
Journal
Journal of Immunological Methods
0022-1759
Publisher
Elsevier
Publication Date
Volume
162
Issue
2
Identifiers
DOI: 10.1016/0022-1759(93)90384-j
Keywords
  • Urokinase-Type Plasminogen Activator (Ec 3.4.21.31)
  • Tissue-Type Plasminogen Activator (Ec 3.4.21.-)
  • Elisa
  • (Human)
Disciplines
  • Biology

Abstract

Abstract ELISA procedures are described for the quantitative analysis of the urokinase-type plasminogen activator (uPA) and of the tissue type PA (tPA). The assays were developed to detect the respective type of PA in cell culture supernatants. TPA can be present as a single-chain or a two-chain protein; uPA as pro-uPA, high or low molecular weight uPA, respectively. In addition, both PAs can be complexed with the plasminogen activator inhibitors PAI-I or PAI-2. Monoclonal antibodies specific for uPA or tPA were selected that recognized the distinct molecular forms of the PAs, even in the presence of fetal calf serum, which is a common — relatively ill-defined — ingredient of cell culture media. The test systems were found to be reliable, easy to perform, and to permit the detection of both types of PA in serum-free and serum-containing cell culture supernatants. Finally, the ELISA — in combination with functional tests — were used to analyse the different PA components in culture supernatants of uPA- or tPA-producing cell lines.

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