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人胸腺白血病相关抗原:分离与部分特性鉴定

Human thymus-leukemia-associated antigen: isolation and partial characterization.

作者信息

Chechik B E, Percy M E, Gelfand E W

出版信息

J Natl Cancer Inst. 1978 Jan;60(1):69-75. doi: 10.1093/jnci/60.1.69.

Abstract

Human thymus-leukemia-associated antigen (HThy-L), a saline-soluble antigen, was previously detected by immunodiffusion (but not on the cell surface) in significant quantity in extracts of normal thymocytes, cells from cultured T-cell lines, and erythrocyte-rosette-positive leukemia blasts. Two species of HThy-L were identified and isolated from normal human thymus tissue after extraction in tris buffer, ammonium sulfate fractionation, acid precipitation of inactive fractions, DEAE-cellulose (DE-52) chromatography, Sephadex G-100 gel filtration, and carboxymethyl-cellulose (CM-52) chromatography; On Sephadex G-100, both HThy-L species had a similar molecular weight (40,000--50,000), but they eluted in different positions on DE-52 and CM-52. Analysis by sodium dodecyl sulfate polyacrylamide gel electrophoresis showed that each of the 2 HThy-L species contained 2 components with molecular weights of approximately 43,000 and 23,000. Further purification of HThy-L on Sephadex G-50 showed that the 43,000-dalton component possessed HThy-L activity.

摘要

人胸腺白血病相关抗原(HThy-L)是一种盐溶性抗原,先前通过免疫扩散法(但不在细胞表面)在正常胸腺细胞提取物、培养的T细胞系细胞以及红细胞玫瑰花结阳性白血病母细胞中大量检测到。在经三羟甲基氨基甲烷缓冲液提取、硫酸铵分级分离、对无活性级分进行酸沉淀、二乙氨基乙基纤维素(DE-52)柱层析、葡聚糖凝胶G-100凝胶过滤以及羧甲基纤维素(CM-52)柱层析后,从正常人胸腺组织中鉴定并分离出两种HThy-L;在葡聚糖凝胶G-100上,两种HThy-L的分子量相似(40,000 - 50,000),但它们在DE-52和CM-52上的洗脱位置不同。十二烷基硫酸钠聚丙烯酰胺凝胶电泳分析表明,两种HThy-L中的每一种都包含分子量约为43,000和23,000的两个组分。在葡聚糖凝胶G-50上对HThy-L进行进一步纯化表明,43,000道尔顿的组分具有HThy-L活性。

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