モノクローナルIgAの作製とその利用

Abstract

We hypothesize that IgA immune complex in mother's milk functions for the prevention of food allergy through oral immune tolerance. To prove this hypothesis and to establish the allergic cure by using IgA immune complex as vaccine, a mass preparation of mouse IgA was performed. We established monoclonal antibody producing hybridomas with mesentery lymph nodes and got three kinds of monoclonal IgA. Their Light chains were all κ and the molecular weight was about 280 kDa, suggesting their dimmer form. The antigen specificity of these antibodies was unidentified, but they did not react with major food allergens. Monoclonal IgA was effectively purified with a Protein L column from the culture supernatant of the hybridoma. Ovalbumin was coupled to the monoclonal IgA with a hetero-bifunctional crosslinking reagent, Sulfosuccinimidyl 6- [3' (2-pyridyldithio) -propionamido] hexanoate. Formation of the pseudo IgA immune complex was confirmed by a sandwich type of Enzyme Linked Immunosorbent Assay with the anti-ovalbumin antibody and the anti-mouse IgA (α) antibody