Mahoney W C, Hermodson M A
J Biol Chem. 1980 Dec 10;255(23):11199-203.
A method employing reverse phase high performance liquid chromatography has been developed for the purification of large denatured peptides. The cyanogen bromide fragments of hemoglobin alpha, beta, and gamma chains (13 to 91 residues in length) could be separated and recovered in 71 to 95% yields using LiChrosorb C-8 columns and a binary solvent system consisting of 0.013 M trifluoroacetic acid in water and a limiting organic solvent. This procedure permits the nondestructive detection of peptides at low wavelength (210 nm) on as little as 50 ng of sample. The peptides are recovered directly by evaporation or lyophilization. An elutropic solvent series for the separation of large and small peptides on reverse phase supports is also described.
已开发出一种采用反相高效液相色谱法来纯化大型变性肽的方法。使用LiChrosorb C - 8柱以及由水中0.013 M三氟乙酸和一种限量有机溶剂组成的二元溶剂系统，可分离并回收血红蛋白α、β和γ链的溴化氰片段（长度为13至91个残基），产率为71%至95%。此程序允许在低波长（210 nm）下对低至50 ng的样品进行无损肽检测。肽可通过蒸发或冻干直接回收。还描述了用于在反相载体上分离大小肽的洗脱溶剂系列。
