Cloning and nucleotide sequence of heavy- and light-chain cDNAs from a creatine-kinase-specific monoclonal antibody

Abstract

Determination of creatine kinase isoenzymes by inhibition assay is a useful tool for the diagnosis and monitoring of myocardial infarction. We have established several mouse hybridoma lines secreting monoclonal antibodies with creatine kinase M-subunit inhibitory capacity. One of the monoclonal antibodies (MAK33) inhibits creatine kinase-MM by 80% without influencing the activity of creatine kinase-MB. A combination of two monoclonal antibodies increased the inhibition of creatine kinase MM up to 99.4%. Poly(A) + RNA of hybridoma cells secreting MAK33 was isolated and used for cloning cDNA of both heavy and light chains of this antibody. Full-length cDNA clones were obtained by hybridization with gamma 1 and kappa constant region cDNA probes. The complete nucleotide sequences from the variable regions including signal peptide and part of the 5'-untranslated regions have been determined.