Quantitative evaluation of stress using metallothionein induced by stress

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It has known that the synthesis of metallothioneins (MTs), metal-binding proteins, is induced by various physical and psychological stressors resulting in increases in MT contents in the liver, blood, and urine. The goal of this study is to establish quantitative evaluation of stress using metallothionein induced under stressful conditions, that is, it is to establish objective evaluation of human stress with high accuracy using MT as an indicator of stress. To establish the evaluation, firstly monoclonal anti MT antibody is to be obtained using human MT, which is used as the antigen, obtained abundantly by a genetic-engineering method. Secondly, MT contents in urine or blood is to be measured by an ELISA method using the anti MT antibody. The results obtained are as follows :1) we successfully obtained the results of MT gene expression by introduction of the human MT II gene into Escherichia coli.2) we obtained abundant refined MT by gel filtration chromatography and high-performance-liquid-chromatography on the crude MT obtained from mass culture of the Escherichia coli.3) we obtained monoclonal anti-MT antibody by cell fusion of mouse myeloma cell and spleen cell of a mouse injected with the human MT II.4) an ELISA method using the monoclonal anti-MT antibody is being developed.