Development of clinical assay method for human chymase

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The original objectives of the project was 1) to develop Westernblot analysis and biochemical assay for plasma human chymase, 2) to determine clinical significance of plasma chymase measurement, 3) to develop RIA method for human chymase to handle massive clinical samples, 4) to compare plasma chymase levels in various cardiovascular diseases, 5) to determine the association between plasma chymase level and various cardiovascular diseases. Among these objectives, objectives 1 and 2 were completed within the first project year, but objectives 3 has been suspended because the iodinated recombinant h-chymase did not bind with the specific antibody. Therefore, we establish a double sandwich ELISA using monoclonal and polyclonal antibodies for recombinant human chymase. Plasma chymase levels significantly increased in systemic inflammatory diseases (pneumonia autoimmune disease and etc.) Mild level of increase was observed in patients with atherosclerosis and ischemic heart disease. There was a significant positive correlation between plasma chymase level and plasma c-reactive protein or fibrinogen. This fact suggests that plasma chymase level may reflect the levels of tissue inflammation. These findings have been reported in Experimental Biology Meeting (Washington DC, USA) and are under considerations for publication in peer review journal. In order to apply this ELISA to clinical use, improvement of the sensitivity will be necessary.