免疫抑制法测定AST线粒体同工酶及其与慢性肝炎、重症肝炎的相关性

目的建立免疫抑制法检测血清AST线粒体同工酶(m-AST)活性,了解血清m-AST活性与肝脏病变程度之间的关系。方法82例正常人、33例慢性肝炎(CH)、19例重型肝炎(FH)病人血清用免疫抑制法和速率法分别测定m-AST和AST总酶活力,算出m-AST/AST比值。结果本法标准曲线y=1.088x-8.889,r=0.999。精密度试验结果为批内CV<3.71%、批间CV<3.91%、回收率96.3～104.6%,平均值100.5%。82例正常血清标本m-AST值和m-AST/AST比值分别为1.1±0.4U/L和5.7±1.8%,33例慢性肝炎血清标本分别为82.6±30.9U/L和26.1±22.5%,19例重型肝炎血清标本分别为152.9±29.5U/L和30.5±25.2%,与正常组比较均有显著性差异(P<0.01)。慢性肝炎和重型肝炎血清标本之间m-AST值和m-AST/AST比值也有显著性差异(P<0.01)。结论免疫抑制法测定血清m-AST方法简单、快速、重复性好,还可间接反映肝细胞损伤的严重程度。     

Immunoelectrophoretic characterization of human mitochondrial aspartate aminotransferase purified by ion exchange and affinity chromatography

Human mitochondrial aspartate aminotransferase (m-ASAT) was prepared for use as an antigen for antibody production and as a standard, i.e. a high degree of purity was demanded. The purification was performed by three ion exchange chromatography steps followed by affinity chromatography on aspartate coupled gel. Four preparations gave specific activities of 230 to 300 U/mg at 37°C. The purity of m-ASAT was assessed by crossed immunoelectrophoresis, which showed that the final preparation did not contain contaminating proteins. Immunization with the purified m-ASAT gave a good antibody response.