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

Abstract

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.