METHODS FOR QUANTITATIVE DETERMINATION OF SERUM ENZYMES USING IMMUNOASSAY AND METHODS FOR DIAGNOSIS OF HEPATIC DISEASE THEREBY

摘要

PURPOSE: A method for quantifying serum enzymes using immunoassay and a method for diagnosis of hepatic disease using the same are provided, thereby accurately diagnosing hepatic disease or progress thereof, and effectively diagnosing chronic hepatic disease. CONSTITUTION: A method for quantifying serum enzyme ALT(alanine aminotransferase) or AST(asphaltate aminotransferase) comprises the steps of: preparing monoclonal antibodies to ALT or AST; constructing hybridoma cell lines producing the monoclonal antibodies; selecting a monoclonal antibody obtained from the hybridoma cell line cultured medium and having improved reactivity with human ALT or AST protein; reacting the selected monoclonal antibody with a blood sample using ELISA to detect serum enzyme ALT or AST which does not bind to an autoantibody, wherein an immunogen used for preparing the monoclonal antibodies is porcine ALT or AST; and the human ALT protein used for selecting a monoclonal antibody is a PCR product using primers having the nucleotide sequences set forth in SEQ ID NO: 1 and SEQ ID NO: 2 and the human AST protein used for selecting a monoclonal antibody is a PCR product using primers having the nucleotide sequences set forth in SEQ ID NO: 3 and SEQ ID NO: 4.