Production of monoclonal antibodies against immunoglobulin heavy chain in common carp (Cyprinus carpio L.)
2006
Vesely, T.(Vyzkumny Ustav Veterinarniho Lekarstvi, Brno (Czech Republic)) E-mail:[email protected] | Reschova, S.(Vyzkumny Ustav Veterinarniho Lekarstvi, Brno (Czech Republic)) | Pokorova, D.(Vyzkumny Ustav Veterinarniho Lekarstvi, Brno (Czech Republic)) | Hulova, J.(Vyzkumny Ustav Veterinarniho Lekarstvi, Brno (Czech Republic)) | Nevorankova, Z.(Vyzkumny Ustav Veterinarniho Lekarstvi, Brno (Czech Republic))
A method for purification of carp serum immunoglobulin (IgM), intended for the production of monoclonal antibodies, is described in the present study. Hybridomas that produce antibodies against IgM heavy chain were selected by ELISA and Western blotting. Ascitic fluids were prepared and tested by the above mentioned methods, followed by their typing. 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 12 fish species, including Oncorhynchus mykiss. Besides Cyprinus carpio, positive results were also found in Carassius auratus, Aristichthys nobilis and Silurus glanis. The sensitivity in common carp was approximately 10 ng/mL.
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