Roos P, Nyberg L, Wide L, Gemzell C
Biochim Biophys Acta. 1975 Oct 20;405(2):363-79.
Two major and two minor components of human luteinizing hormone (lutropin) were isolated from whole frozen pituitaries by a procedure involving extraction of homogenized pituitaries, (NH4)2SO4 fractionation, chromatography on DEAE-cellulose, Sephadex G-100, and SE-Sephadex C-50 and electrophoresis in polyacrylamide gel. The isolation procedure was monitored by both bioassays and radioimmunoassays. Contamination of the final products by other pituitary hormone activities was very low. The four lutropin components were all homogeneous by polyacrylamide gel electrophoresis (a sieving medium) and by free zone electrophoresis (a non-sieving medium). No heterogeneity was observed when the components were studied in the ultracentrifuge by sedimentation-equilibrium technique. The molecular weights of the components were in the range of 34 000-40 000. Sedimentation velocity experiments with the two major components revealed in each case one boundary with S20,W values of 3.2 S and 3.5 S. Further evidence for the homogeneity of the components was the observation of only one precipitin line for each component upon immunodiffusion against a rabbit anti-human lutropin serum. Amino acid and carbohydrate analyses indicated close similarity among the four components. From the analysis data the molecular weights of the components were calculated to be 31 000-33 000.
通过一系列步骤从冷冻的人垂体中分离出人类促黄体生成素（促性腺激素）的两种主要成分和两种次要成分，这些步骤包括对匀浆垂体进行提取、硫酸铵分级分离、在二乙氨基乙基纤维素、葡聚糖凝胶G - 100和强阳离子交换葡聚糖凝胶C - 50上进行色谱分离以及在聚丙烯酰胺凝胶中进行电泳。通过生物测定和放射免疫测定对分离过程进行监测。最终产物受其他垂体激素活性的污染非常低。这四种促黄体生成素成分通过聚丙烯酰胺凝胶电泳（一种筛分介质）和自由区电泳（一种非筛分介质）均表现为均一。当通过沉降平衡技术在超速离心机中研究这些成分时，未观察到异质性。这些成分的分子量在34000 - 40000范围内。对两种主要成分进行沉降速度实验，在每种情况下均显示出一个边界，其S20,W值分别为3.2 S和3.5 S。这些成分均一性的进一步证据是，在用兔抗人促黄体生成素血清进行免疫扩散时，每种成分仅观察到一条沉淀线。氨基酸和碳水化合物分析表明这四种成分之间非常相似。根据分析数据计算出这些成分的分子量为31000 - 33000。
