Development and in vitro characterization of a new immunoassay of cardiac troponin T.
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%).
Katus
H A
HA
Innere Medizin III, University of Heidelberg, F.R.G.
Looser
S
S
Hallermayer
K
K
Remppis
A
A
Scheffold
T
T
Borgya
A
A
Essig
U
U
Geuss
U
U
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D001426:Bacterial Proteins; D002417:Cattle; D006801:Humans; D007124:Immunoenzyme Techniques / Q000592:standards / Q000706:statistics & numerical data; D008969:Molecular Sequence Data; D009132:Muscles / Q000737:chemistry*; D009206:Myocardium / Q000737:chemistry*; D019809:Streptavidin; D014336:Troponin / Q000032:analysis* / Q000097:blood / Q000737:chemistry; D020107:Troponin T
D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D014336:Troponin; D020107:Troponin T; D019809:Streptavidin