Sweet F, Szabados L
Steroids. 1977 Jan;29(1):127-44. doi: 10.1016/0039-128x(77)90115-5.
Synthesis of an estrogen affinity adsorbent containing a disulfide linkage between the steroid and stationary matrix permitted facile purification of high affinity estrogen binding proteins. Following affinity chromatography of either antibody directed against estrone 17-carboxymethyloxime - bovine serum albumin or immature calf uterine cytoplasmic estrogen receptor proteins, the specifically bound mercaptoethanol. Crude antibody and uterine cytosol was 10(-3) to 10(-2)M cystamine (S-S) to block SH-containing proteins, in order to protect the adsorbent against protein-mediated S-S in equilibrium SH exchange. Cystamine was found to markedly stabilize crude cytosol receptor protein by 200-300% compared with preparations obtained under ordinary conditions. Disulfide affinity adsorbents are versatile in that they can be used either under conventional conditions of specific protein recovery, or with 2-mercaptoethanol which removes the ligand and bound protein from the stationary matrix quantitatively.
合成一种在类固醇与固定基质之间含有二硫键的雌激素亲和吸附剂，可轻松纯化高亲和力雌激素结合蛋白。在用针对雌酮17 - 羧甲基肟 - 牛血清白蛋白的抗体或未成熟小牛子宫细胞质雌激素受体蛋白进行亲和层析后，特异性结合的巯基乙醇。粗制抗体和子宫胞液用10(-3)至10(-2)M胱胺（S - S）来阻断含SH的蛋白质，以保护吸附剂免受蛋白质介导的S - S在平衡时的SH交换。与在普通条件下获得的制剂相比，发现胱胺可使粗制胞液受体蛋白显著稳定200 - 300%。二硫键亲和吸附剂用途广泛，因为它们既可以在特定蛋白质回收的常规条件下使用，也可以与2 - 巯基乙醇一起使用，后者可定量地从固定基质中去除配体和结合蛋白。
