Analysis of the antigen epitopes of infectious pathogens in laboratory animals using peptide tips

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We selected mouse hepatitis virus, Sendai virus, Mycoplasma plumonis, and hantavirus from the aspect of the frequency and danger upon infectious accidents. We determined antigenic epitopes by incubating polyclonal anti-sera with peptide tips produced according to the amino acid sequences reported previously elsewhere. Further, we tried to develop a preliminary ELISA kit using antigenic epitopes that we have determined. With respect to mouse hepatitis virus and Sendai virus, we succeeded to develop an ELISA system, which was more sensitive or equal to the commercially available ELISA system (paper in preparation). With respect to hantavirus, we succeeded to determine an antigenic epitope, EDVNGIRK (amino acid 166-173), to monoclonal antibody E5/G6, which recognizes nuclear protein of hantavirus. With respect to Mycoplasma plumonis, we first searched antigenic protein from the components of the bacteria, since there are a lot of proteins composing of bacteria. We found a protein showing a homology to P46 protein, which had been reported as an antigenic protein in other Mycoplasmas and named it P46L. We produced recombinant P46L protein conjugated with glutathione S-transferase and made an ELISA system with purified P46L. This ELISA system was more sensitive than the commercially available ELISA system.We further applied this method to the determination of antigenic epitopes of West Nile virus and SARS virus and succeeded it. We furthermore succeeded to apply this method to not only determining antigenic epitopes of infectious pathogens but also determining protein-protein interaction site such as binding sits of FAS-antigen and angiotensin II.