ANALYSIS OF EFFECTIVENESS OF USING IgY FROM CHICKEN IN SANDWICH METHOD OF HBsAg TESTING

Abstract

Study antigen-binding ability of polyclonal antibodies (PCA) of chicken compared with monoclonal -antibodies (MCA) of mice in the model of interaction with HBsAg. Mice MCA 18C8 and MKA F3/F4 (IgG) were used, effective in enzyme immunoassay sandwich method of HBsAg determination (with a minimal detection dose of 0.017 ng/ml), and affinity purified anti-HBsAg PCA of chicken (IgY), obtained from 2 immunized birds (PCA No. 1 and PCA No. 2). The ability of antibodies to bind HBsAg was evaluated by analytical sensitivity (slope of binding curve) of solid-phase enzyme immunoassay system using mice MCA and chicken PCA. PCA No. 2 has provided a statistically significant 40% increase of analytical sensitivity, compared with <<standard immobilized MCA 18C8, in model experiments of binding of peroxidase-labeled HBsAg. However, transition from model experiments to use of PKA No. 1 and PKA No. 2 in sandwich method of determination of HBsAg instead of immobilized MCA 18C8 or detecting MCA F3/F4 in all the cases, on the contrary, resulted in a decrease of analytical sensitivity. A lower flexibility of chicken PCA was assumed to be able to impede bivalent interaction in sandwich method, resulting in formation of less stable imminne complexes. Without Challenging value of IgY for the creation of immunochemical diagnostic methods, these facts and assumptions indicate a necessity of a deeper elucidation of the best ar- eas of their application.