Dobos M, Burger H G, Hearn M T, Morgan F J
Mol Cell Endocrinol. 1983 Aug;31(2-3):187-98. doi: 10.1016/0303-7207(83)90148-x.
This report describes the application of high performance liquid chromatography (HPLC) and associated techniques to the isolation of inhibin from ovine follicular fluid (oFF). Two modes of HPLC have been assessed. High speed gel permeation chromatography on a Waters I-125 support showed that at pH 7.0 inhibin bioactivity was associated with an approx. 80000 mol. wt protein fraction which co-eluted with the bulk of the protein in oFF and gave a yield of 30-44% of the applied activity. Reversed-phase HPLC (RP-HPLC) of a peptide-rich fraction extracted from oFF was also investigated. Using analytical liquid chromatography on octadecylsilylsilica (ODS-silica), two distinct inhibin bioactive fractions were obtained, with a resultant purification of 23-fold and 3-fold respectively and an overall recovery of 8.5%. These results demonstrate that RP-HPLC may be employed as a direct and rapid technique for the isolation of inhibin from oFF.
本报告描述了高效液相色谱（HPLC）及相关技术在从绵羊卵泡液（oFF）中分离抑制素方面的应用。评估了两种HPLC模式。在Waters I - 125载体上进行的高速凝胶渗透色谱显示，在pH 7.0时，抑制素生物活性与一种分子量约为80000的蛋白质组分相关，该组分与oFF中的大部分蛋白质共洗脱，产率为所施加活性的30 - 44%。还研究了从oFF中提取的富含肽的组分的反相HPLC（RP - HPLC）。使用十八烷基硅烷硅胶（ODS - 硅胶）进行分析液相色谱，得到了两个不同的抑制素生物活性组分，纯化倍数分别为23倍和3倍，总回收率为8.5%。这些结果表明，RP - HPLC可作为一种直接且快速的技术用于从oFF中分离抑制素。
