Production and characterization of monoclonal antibodies to portunus trituberculatus hemocyanin

Abstract

Murine monoclonal antibodies to hemocyanin of the crab, Portunus trituberculatus was derived from somatic cell fusion between Balb/c immune splenocytes and myeloma. Six hybridoma lines were selected and cloned, each of them sectrets IgG 1 and exhibits titres around those of 500-1,000 fold diluted antisera. One of six monoclonal antibodies were extensively characterized and used for studying cognate antigen. Immunochemical relationship with arthropodan hemocyanin and effect on pseudo-peroxidatic activity were studied with conventional antiserum and monoclonal antibody. Six monoclonal antibodies were shown to be inhibitory in peroxidatic reaction of Portunus hemocyanin, anti-Portunus hemocyanin monoclonal antibody (POTE53) and serum was cross-reactive with keyhole limpet hemoyanin in competititive radioimmune assay whereas antisera of Portunus hemocyanin and keyhole limpet hemocyanin were not cross-reactive in double immunodiffusion.