Direct measurement of creatine kinase-MB activity in serum after extraction with a monoclonal antibody specific to the MB isoenzyme.
Fusion of splenocytes from A/J mice immunized by creatine kinase (EC 2.7.3.2)-MB isoenzyme (CK-MB) with SP2/0-Ag14 myeloma cell line generated hybridomas producing monoclonal antibodies specific to CK-MB. One of these monoclonal antibodies ("Conan-MB") was used to develop a direct assay for CK-MB activity. In the assay, serum is incubated for 30 min at room temperature with "Conan-MB" immobilized on latex beads. The beads are then washed, and CK-MB activity bound to the antibody is measured after incubation with CK enzyme reagent for 30 min at 37 degrees C. Results with the assay correlated well (r = 0.997) with those for CK-MB concentration as measured by a two-site immunoassay. Neither CK-MM, CK-BB, mitochondrial CK, nor a hemolysate of erythrocytes interfered. Use of this unique monoclonal antibody allows rapid, precise, and direct determination of CK-MB activity.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification; D000918:Antibody Specificity; D001201:Ascites / Q000276:immunology; D003402:Creatine Kinase / Q000097:blood* / Q000276:immunology; D004587:Electrophoresis, Agar Gel; D005260:Female; D006801:Humans; D007118:Immunoassay / Q000379:methods; D007527:Isoenzymes; D007840:Latex; D051379:Mice; D008805:Mice, Inbred A
D000911:Antibodies, Monoclonal; D007527:Isoenzymes; D007840:Latex; D003402:Creatine Kinase