Assay of Human Tissue-Type Plasminogen Activator (t-PA) with an Enzyme-Linked Immunosorbent Assay (ELISA) Based on Three Murine Monoclonal Antibodies to t-PA

Paul Holvoet; Hugo Cleemput; Désiré Collen

doi:10.1055/s-0038-1660097

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 1985; 54(03): 684-687
DOI: 10.1055/s-0038-1660097

Original Article

Schattauer GmbH Stuttgart

Assay of Human Tissue-Type Plasminogen Activator (t-PA) with an Enzyme-Linked Immunosorbent Assay (ELISA) Based on Three Murine Monoclonal Antibodies to t-PA

Paul Holvoet

The Center for Thrombosis and Vascular Research, University of Leuven, Belgium

,

Hugo Cleemput

The Center for Thrombosis and Vascular Research, University of Leuven, Belgium

,

Désiré Collen

The Center for Thrombosis and Vascular Research, University of Leuven, Belgium

› Institutsangaben

Abstract

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Summary

An enzyme-linked immunosorbent assay (ELISA) for the measurement of human tissue-type plasminogen activator (t-PA) was developed. Microtiter plates were coated with a mixture of two monoclonal antibodies and bound t-PA was quantitated with a third monoclonal antibody linked to peroxidase. The lower limit of sensitivity of the assay was 0.2 ng of t-PA per ml. The concentration of antigen in citrated plasma of human subjects was found to be 3.4 ± 0.8 ng/ml. The assay had a good reproducibility with values of 3.8, 6.5 and 4.9 percent respectively for the intra-, inter-assay and inter-dilution variation coefficients. The results of the ELISA assay on plasma samples from patients during thrombolytic therapy with t-PA correlated very well, over a wide concentration range, with those obtained with a previously described two-site immuno-radiometric assay (r = 0.96). This ELISA with monoclonal antibodies constitutes a stable and reproducible set of reagents for the measurement of t-PA antigen in biological fluids, avoiding the disadvantages of the use of radioisotopes and of polyclonal antibodies.

Keywords

ELISA - Tissue-type plasminogen activator - Monoclonal antibodies

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Referenzen

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