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Clinical Chemistry 38: 386-393, 1992;
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Clinical Chemistry, Vol 38, 386-393, Copyright © 1992 by American Association for Clinical Chemistry

Development and in vitro characterization of a new immunoassay of cardiac troponin T

HA Katus, S Looser, K Hallermayer, A Remppis, T Scheffold, A Borgya, U Essig and U Geuss
Innere Medizin III, University of Heidelberg, F.R.G.

We describe a new one-step enzyme immunoassay of troponin T that uses two specific monoclonal antibodies and streptavidin-coated tubes as the solid phase. The monoclonal antibodies were obtained by conventional hybridoma technology, with human troponin T as antigen. The identity of the cardiac troponin T antigen was confirmed by analysis of the amino acid composition and by partial sequence analysis. The specificity of the monoclonal antibodies for cardiac troponin T was proved by immunoblot analysis and displacement curves. The capture antibody is labeled with biotin. The second antibody is conjugated to horseradish peroxidase (EC 1.11.1.7). The assay [Enzymun-TestR system (Boehringer Mannheim GmbH)] is performed in only 90 min at room temperature; the measuring range for troponin T is 0.1 to 15 micrograms/L. The assay shows excellent between-run precision (CV = 3.3-4.9%).


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