title: Immunochemiluminometric assay of creatine kinase MB with a monoclonal antibody to the MB isoenzyme.
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 susceptible 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 microgram 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 micrograms/L. Results for serum samples by the new assay correlated well (r = 0.94) with those by Corning electrophoretic and the Hybritech 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.
authors.forename: U
authors.initials: U
authors.lastname: Piran
authors.forename: D W
authors.initials: DW
authors.lastname: Kohn
authors.forename: L S
authors.initials: LS
authors.lastname: Uretsky
authors.forename: D
authors.initials: D
authors.lastname: Bernier
authors.forename: E H
authors.initials: EH
authors.lastname: Barlow
authors.forename: C A
authors.initials: CA
authors.lastname: Niswander
authors.forename: M
authors.initials: M
authors.lastname: Stastny
mesh_terms: D000166:Acridines; D000911:Antibodies, Monoclonal; D003402:Creatine Kinase / Q000097:blood*; D003429:Cross Reactions; D004586:Electrophoresis; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007118:Immunoassay / Q000379:methods*; D007124:Immunoenzyme Techniques; D007202:Indicators and Reagents; D007527:Isoenzymes; D008163:Luminescent Measurements; D008280:Magnetics; D011786:Quality Control; D011933:Reagent Kits, Diagnostic
chemical_list: D000166:Acridines; D000911:Antibodies, Monoclonal; D007202:Indicators and Reagents; D007527:Isoenzymes; D011933:Reagent Kits, Diagnostic; D003402:Creatine Kinase