Brent T P, von Wronski M, Pegram C N, Bigner D D
Department of Biochemical and Clinical Pharmacology, St. Jude Children's Research Hospital, Memphis, Tennessee 38101.
Cancer Res. 1990 Jan 1;50(1):58-61.
As well as repairing mutagenic lesions induced by simple methylating agents, O6-alkylguanine-DNA alkyltransferase repairs precursors of cytotoxic interstrand cross-links induced by chloroethylating anticancer drugs. Moreover, levels of the transferase correlate with cellular resistance to these agents. Thus far, the human transferase has not been highly purified. In our quest to obtain the homogeneous protein we have produced four stable cloned hybridomas that secrete monoclonal antibodies against the alkyltransferase from human lymphoblasts. The specificity of these monoclonals was established by immunoblot analysis and immunoprecipitation. All these antibodies recognized the alkyltransferase only after its denaturation by sodium dodecyl sulfate. One of them, designated 19.2, was used in immunoaffinity chromatography to obtain the pure protein.
O6-烷基鸟嘌呤-DNA烷基转移酶不仅能修复由简单甲基化剂诱导的诱变损伤，还能修复由氯乙基化抗癌药物诱导的细胞毒性链间交联的前体。此外，该转移酶的水平与细胞对这些药物的抗性相关。到目前为止，人源转移酶尚未得到高度纯化。在我们获取纯蛋白的过程中，我们制备了四种稳定的克隆杂交瘤，它们分泌针对人淋巴母细胞烷基转移酶的单克隆抗体。通过免疫印迹分析和免疫沉淀确定了这些单克隆抗体的特异性。所有这些抗体只有在十二烷基硫酸钠使其变性后才能识别烷基转移酶。其中一种命名为19.2的抗体被用于免疫亲和色谱以获得纯蛋白。
