High-resolution reversed-phase high-performance liquid chromatography of peptides and proteins.

摘要: Publisher Summary Reversed-phase high-performance liquid chromatography (RP-HPLC) has become a commonly used method for the analysis and purification of peptides proteins. The extraordinary popularity RP-HPLC can be attributed to number factors, including excellent resolution that achieved closely related as well structurally disparate substances under large variety chromatographic conditions; experimental ease, with which selectivity manipulated through changes in mobile phase composition; generally high recoveries, even at ultramicroanalytical levels; reproducibility repetitive separations carried out over long periods time, due part stability various sorbents many productivity terms cost parameters; potential is only now being addressed evaluation different physicochemical aspects solute-eluent or solute-hydrophobic sorbent interactions assessment their structural consequences from data. system usually comprises an n-alkylsilica-based sorbent, proteins are eluted, gradients increasing concentration organic solvent, such acetonitrile, containing ionic modifier, e.g., trifluoroacetic acid (TFA). With modern instrumentation columns, complex mixtures separated low picomolar amounts resolved components collected. Separations easily manipulated, by changing gradient slope, temperature, solvent composition. technique equally applicable enzymatically derived also synthetically peptides.