Development and performance of an enzyme immunoassay to detect creatine kinase isoenzyme MB activity using anti-mitochondrial creatine kinase monoclonal antibodies

Scand J Clin Lab Invest. 2009;69(6):687-95. doi: 10.3109/00365510902981171.

Abstract

Objective: The MB fraction of creatine kinase (CK-MB) has long been used as a cardiac marker. It is known that the CK-MB immunoinhibition method lacks selectivity and accuracy, because the appearance of macro CK type 2, corresponding to mitochondrial creatine kinase (MtCK) in some patient serum may render CK-MB activity measured by conventional method abnormally high. Thus, to improve the specificity and accuracy of the CK-MB assay, we developed two types of monoclonal anti-MtCK antibodies against sarcomeric MtCK and ubiquitous MtCK, and present herein the performance of a new method using these antibodies.

Material and methods: The performance of our test for detecting CK-MB activity was compared with other methods, and the range of CK-MB activities in normal human serum was investigated.

Results: The two types of monoclonal antibodies developed by us were isoenzyme-specific to sMtCK or uMtCK. The correlation coefficients of our method and conventional method to electrophoresis were 0.973 and 0.873, respectively. The mean CK-MB activity in normal human serum by our method and the conventional method was 2.4 and 11.7 U/L, respectively. Thus, our data indicated that about 80% of CK-MB activity, determined using the conventional method, seems to correspond to the MtCK activity.

Conclusion: Our method is novel in offering higher accuracy of measuring true CK-MB contents in human serum as compared to the conventional method. The possibility of accurately estimating CK-MB activity by our method which can inhibit MtCKs in healthy person and patient serum is likely to bring a break-through in clinical diagnostics.

MeSH terms

  • Antibodies, Monoclonal / pharmacology*
  • Antibody Specificity / drug effects
  • Binding Sites, Antibody
  • Creatine Kinase, BB Form / antagonists & inhibitors
  • Creatine Kinase, BB Form / blood
  • Creatine Kinase, MB Form / antagonists & inhibitors
  • Creatine Kinase, MB Form / blood*
  • Creatine Kinase, Mitochondrial Form / antagonists & inhibitors
  • Creatine Kinase, Mitochondrial Form / blood
  • Creatine Kinase, Mitochondrial Form / immunology*
  • Electrophoresis
  • Health
  • Humans
  • Immunoenzyme Techniques / methods*
  • Immunoenzyme Techniques / standards*
  • Isoenzymes / antagonists & inhibitors
  • Isoenzymes / blood
  • Membranes, Artificial
  • Molecular Weight
  • Reference Values

Substances

  • Antibodies, Monoclonal
  • Isoenzymes
  • Membranes, Artificial
  • Creatine Kinase, BB Form
  • Creatine Kinase, MB Form
  • Creatine Kinase, Mitochondrial Form