Immunochemiluminometric assay of creatine kinase MB with a monoclonal antibody to the MB isoenzyme.
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.
Piran
U
U
Kohn
D W
DW
Uretsky
L S
LS
Bernier
D
D
Barlow
E H
EH
Niswander
C A
CA
Stastny
M
M
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
D000166:Acridines; D000911:Antibodies, Monoclonal; D007202:Indicators and Reagents; D007527:Isoenzymes; D011933:Reagent Kits, Diagnostic; D003402:Creatine Kinase