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多种疾病能导致血清核糖核酸酶(RNase)活性升高。目前血清核糖核酸酶浓度的测定采用酶学方法。但该法不能区分核糖核酸酶的来源。本文用肝素亲和层析等一系列技术来纯化人肝核糖核酸酶,并用此制剂首次建立了一种敏感、特异而且重复性好的人肝RNase放射免疫测定法。人肝RNase的纯化及其特征通过缓冲液抽提、酸分离、磷酸纤维素柱层析、Sephadex G-150和Sephadex G-100凝胶层析、聚鸟嘌岭核苷酸亲和层析和肝素亲和层析等步骤,从人肝脏组织中提取并纯化了核糖核酸酶。纯化后的人肝核糖核酸酶在SDS聚丙烯酰胺凝胶电泳上呈单一的狭窄区带;在pH4.3聚丙烯酰胺凝胶电泳上呈单一的,但略为扩散的区带。酶活性和蛋白区带相吻合。该酶由SDS聚丙烯酰胺电泳测
A variety of diseases can lead to elevated serum RNase activity. At present, the concentration of serum ribonucleases is determined enzymatically. However, this method can not distinguish the source of ribonucleases. In this paper, a series of techniques such as heparin affinity chromatography to purify human liver RNase, and for the first time with this preparation to establish a sensitive, specific and reproducible human liver RNase radioimmunoassay. Purification and characterization of human liver RNase was performed by buffer extraction, acid separation, phosphocellulose column chromatography, Sephadex G-150 and Sephadex G-100 gel chromatography, polyGUIDA and Heparin affinity chromatography and other steps, extracted from human liver tissue and purified ribonuclease. The purified human liver RNase showed a single narrow zone on SDS polyacrylamide gel electrophoresis and a single but slightly diffused zone on pH4.3 polyacrylamide gel electrophoresis. Enzyme activity and protein band coincide. The enzyme was electrophoresed on SDS polyacrylamide