Analysis of immune-reactive components of ayu serum using monoclonal antibody

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The subject of the present study was to make monoclonal antibodies against serum components especially main immune reactive components of ayu, Plecoglossus altivelis, to detect and quantitatively determine those components.Trials to make monoclonal antibodies were done repeatedly by screening and cloning hybridomas of established mouse myeloma cells and lymphocytes obtained from the spleen of mice immunized with ayu serum by means of immune blotting and enzyme-linked immunosorbent assay (ELISA) with polystyrene beads sensitized with ayu serum using anti-mouse IgG labeled with peroxydase. To make anti-ayu antibody monoclonal antibody, ELISA using the serum of ayu immunized with Vibrio anguillarum and polystyrene beads sensitized with V.anguillarum lipopolysaccharide. Detection was done on the serum and homogenates of the kidney, spleen and liver of immunized and unimmunized ayu. Detection was dine also on eggs and fry of ayu obtained from a hatchery.Totally twelve monoclonal antibodies ("A"-"L") were obtained. Monoclonal antibody "A" was considered to be that against ayu antibody. Antigens "a"-'1", which corresponded to monoclonal antibodies "A"-"L", were detected in the serum and the homogenates of the kidney, spleen and liver and the fries but only "b"-"e" and "1" were detected in the eggs. Concentration of the "a" was higher in the immunized ayu than that in the unimmumized ayu, but the concentration of other antigens did not increasae by immunization of ayu.