Chechik B E, Madapallimattam A, Gelfand E W
J Natl Cancer Inst. 1979 Mar;62(3):465-70. doi: 10.1093/jnci/62.3.465.
A competitive radioimmunoprecipitation method was developed for the quantitation of human thymus-leukemia-associated antigen (HThy-L) isolated from human thymus tissue. Antisera to the antigen were raised by immunization of rabbits with purified fractions of HThy-L. The antigen was labeled with 125l by means of a modification of the lactoperoxidase technique and subjected to Sephadex G-100 gel filtration. Analysis of fractions eluted from this column by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed two labeled components with apparent molecular weights of 45,000 and 25,000 daltons, respectively. Immunoprecipitation with anti-HThy-L antiserum demonstrated directly that the 45,000-dalton component carried HThy-L antigenicity. This fraction served as the source of labeled antigen in a competitive radioimmunoassay that permitted the detection of approximately 1 ng HThy-L. With the use of this assay, we confirmed quantitatively that the highest amounts of HThy-L were found in extracts of thymocytes, normal thymus tissues, and lymphoblasts for T-cell lines.
已开发出一种竞争性放射免疫沉淀法，用于定量从人胸腺组织中分离出的人胸腺白血病相关抗原（HThy-L）。通过用纯化的HThy-L组分免疫兔子来制备针对该抗原的抗血清。借助改良的乳过氧化物酶技术用125I标记该抗原，并进行Sephadex G-100凝胶过滤。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳对从该柱洗脱的级分进行分析，发现有两个标记组分，其表观分子量分别为45,000和25,000道尔顿。用抗HThy-L抗血清进行免疫沉淀直接证明，45,000道尔顿的组分具有HThy-L抗原性。该级分用作竞争性放射免疫测定中标记抗原的来源，该测定可检测到约1 ng的HThy-L。使用该测定法，我们定量证实，在胸腺细胞、正常胸腺组织和T细胞系的淋巴母细胞提取物中发现的HThy-L含量最高。
