Immunochemiluminometric assay of creatine kinase MB with a monoclonal antibody to the MB isoenzyme

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Abstract

Previous two-site immunometric assays for creatine kinase (CK; EC 2.7.3.2) MB isoenzyme have been based on formation of a 'sandwich' complex involving CK-MB and antibodies that recognize the CK-MM and the CK-BB isoenzymes. Single-incubation model assays of CK-MB with these antibodies were suscpetible to interferences by CK-MM and CK-BB. We produced two anti-CK-MB monoclonal antibodies and studied their suitability for two-site assays. Both antibodies were compatible with anti-CK-MM and anti-CK-BB, but not with each other. Using anti-CK-MB as the tracer antibody eliminated the interference by both CK-MM and CK-BB. Labeling anti-CK-MB with acridinium ester and immobilizing anti-CK-BB on paramagnetic particles, we developed a rapid and highly sensitive chemiluminescent/magnetic separation CK-MB assay. As little as 1 μg of CK-MB per liter was detectable after 10- or 30-min incubation at room temperature, and the standard curve was linear up to 400 μg/L. Results for serum samples by the new assay correlated well (r = 0.94) with those by Corning electrophoretic and the Hyberitech Tandem-E immunoenzymometric CK-MB methods. Sera containing macro CK-1 or high concentrations of CK-MM and CK-BB did not interfere. The combined advantages of a more-specific antibody, paramagnetic solid phase, and chemiluminescent label endow this two-site CK-MB assay with performance characteristics and ease of use superior to those of previous assays.

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Piran, U., Kohn, D. W., Uretsky, S., Bernier, D., Barlow, E. H., Niswander, C. A., & Stastny, M. (1987). Immunochemiluminometric assay of creatine kinase MB with a monoclonal antibody to the MB isoenzyme. Clinical Chemistry, 33(9), 1517–1520. https://doi.org/10.1093/clinchem/33.9.1517

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