Applications of preparative high-performance liquid chromatography to the separation and purification of peptides and proteins

The basic differences between the problems encountered in the development of the analytical and preparative applications of high-performance liquid chromatography are analyzed and the principles of optimization of a preparative separation outlined. The application of these principles to the purification of peptides and proteins are discussed with emphasis on the specific problems raised by the separation of high-molecular-mass compounds. The relative advantages and inconveniences of displacement and overloaded elution are presented. The applications of the various retention modes of liquid chromatography (ion exchange, reversed-phase adsorption, hydrophobic interactions, size exclusion and bioaffinity) to the separation of various peptide and protein mixtures and/or to the extraction and purification of some of these compounds are reviewed. Separation performance reported in the literature are summarized in table form.