Assay of creatine kinase isoenzyme MB in serum with time-resolved immunofluorometry.
We describe the first time-resolved immunofluorometric assay for creatine kinase (EC 2.7.3.2) isoenzyme MB (CK-MB) in serum. The assay is based on the formation of the complex: solid-phase anti-CK-MB-CK-MB-biotinylated anti-CK-BB-streptavidin-BCPDA-Eu3+, where anti-CK-MB and anti-CK-BB are monoclonal antibodies against the CK isoenzymes MB and BB, respectively, and BCPDA is the europium chelator 4,7-bis(chlorosulfophenyl)-1,10-phenanthroline-2,9-dicarboxylic acid. The solid-phase complex is fluorescent and is measured on the dry solid-phase (microtiter well) in a specially designed time-resolved fluorometer that uses laser excitation. The assay requires 25 microL of serum and is not affected by the presence of either CK-MM (up to 5000 micrograms/L) or CK-BB (up to 1000 micrograms/L) in the sample. Precision and accuracy indices for the assay were satisfactory.
Department of Clinical Biochemistry, Toronto Western Hospital, Ontario, Canada.
E S
ES
Lianidou
T K
TK
Christopoulos
E P
EP
Diamandis
D000911:Antibodies, Monoclonal; D003402:Creatine Kinase / Q000097:blood*; D005063:Europium; D015200:Fluoroimmunoassay; D006801:Humans; D007527:Isoenzymes
D000911:Antibodies, Monoclonal; D007527:Isoenzymes; D005063:Europium; D003402:Creatine Kinase