ISOLATION AND CHARACTERIZATION OF 5T4, A TUMOR-ASSOCIATED ANTIGEN
ISOLATION AND CHARACTERIZATION OF 5T4, A TUMOR-ASSOCIATED ANTIGEN
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DOI：
10.1002/ijc.2910450132
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发表时间：
1990-01-15
影响因子：
6.4
通讯作者：
STERN, PL
中科院分区：
文献类型：
--
作者：
HOLE, N;STERN, PL
单克隆抗体 (MAb) 5T4 定义了一种人滋养层抗原标记物，在正常成人组织中具有有限的表达模式，但该抗原在多种癌症中表达。 5T4 抗原分子的纯化从术语合体滋养层开始描述，通过凝集素和免疫亲和层析以及凝胶过滤的组合，提供高达 10,00 倍的纯化，产率 70%。该抗原由非缔合糖蛋白分子携带，在 SDS-PAGE 上表观分子量为 72 kDa，pI 为中性。 N-聚糖酶去除 N-连接糖揭示了 42 kDa 的核心蛋白。用切割 O-连接糖的酶处理不会显着改变分子大小。天然 5T4 分子对蛋白水解具有很强的抵抗力，直到 N 连接糖被去除或糖蛋白变性和还原。通过这些方法产生的糖肽将适用于氨基酸测序。
The monoclonal antibody (MAb) 5T4 defines a human trophoblast antigen marker with a restricted pattern of expression in normal adult tissues but this antigen is expressed on a variety of carcinomas. The purification of 5T4 antigenic molecules is described from term syncytiotrophoblast by a combination of lectin- and immunoaffinity chromatography and gel filtration giving up to 10,00-fold purification with 70% yield. The antigen is carried by non-associated glycoprotein molecules with an apparent molecular weight of 72 kDa on SDS-PAGE and a neutral pI. Removal of N-linked sugars by N-glycanase reveals a core protein of 42 kDa. Treatment with enzymes that cleave O-linked sugars does not substantially alter the molecular size. The native 5T4 molecules are very resistant to proteolysis until the N-linked sugars are removed or the glycoprotein is denatured and reduced. Glycopeptides generated by these approaches will be suitable for amino acid sequencing.