Production of monoclonal antibodies against immunoglobulin heavy chain in common carp (Cyprinus carpio L.)

Abstract

A method for purification of carp serum immunoglobulin (IgM), intended for the production of monoclonal antibodies, was described in the present study. Hybridomas that produce antibodies against IgM heavy chain were selected by ELISA method and Western blotting. Ascitic fluids were prepared and tested by the above mentioned methods, and their typing followed. Monoclonal antibody with the highest titre of antibodies against carp immunoglobulin was selected for conjugation with horseradish peroxidase. Specificity of conjugated monoclonal antibody was tested in a panel of various fish species sera. Cross-reactivity was not detected in rainbow trout (Oncorhynchus mykiss) and eleven other fish species. Besides common carp, positive results were also found in goldfish (Carassius auratus) and bighead carp (Aristichthys nobilis), that are members of Cyprinidae family. Among fish other than Cyprinidae, positive results were also detected in sheatfish (Silurus glanis). The sensitivity in common carp was approximately 10 ng/ml.