Ozawa M, Muramatsu T, Solter D
Cell Differ. 1985 May;16(3):169-73. doi: 10.1016/0045-6039(85)90514-7.
Molecules carrying SSEA-1 were isolated from [3H]galactose-labeled embryonal carcinoma cells by detergent solubilization followed by indirect immunoprecipitation. The antigenic molecules were degraded by extensive pronase digestion or mild alkaline treatment, and the majority of the products thus formed were so large as to be excluded from a column of Sephadex G-50. Therefore, the major carbohydrate constituent of the antigenic molecule was embryoglycan, the glycoprotein-bound large glycan in early embryonic cells. Furthermore, the binding of isolated embryoglycan with anti-SSEA-1 was directly shown by a modified Farr's assay. From these results we concluded that SSEA-1 determinant was carried by the large glycan.
通过去污剂溶解,随后进行间接免疫沉淀,从[³H]半乳糖标记的胚胎癌细胞中分离出携带阶段特异性胚胎抗原-1(SSEA-1)的分子。抗原分子经大量蛋白酶消化或温和碱性处理后会降解,由此形成的大多数产物都很大,以至于无法通过葡聚糖凝胶G-50柱。因此,抗原分子的主要碳水化合物成分是胚胎聚糖,即早期胚胎细胞中糖蛋白结合的大聚糖。此外,通过改良的法尔氏试验直接证明了分离出的胚胎聚糖与抗SSEA-1的结合。从这些结果我们得出结论,SSEA-1决定簇由大聚糖携带。