ABSTRACT
The efficient isolation of peptides has become increasingly important for an ever-widening range of research disciplines in recent years and high-performance liquid chromatography (HPLC) has shown immense versatility in the separation and purification of peptides from a great variety of sources. The three main modes of HPLC used for peptide separations utilize differences in peptide size, net charge or hydrophobicity. A computer software program, ProDigest-LC, has been developed to assist scientists in devising methods for the analytical separation and purification of biologically active peptides and peptide fragments from enzymatic and chemical digests of proteins by size-exclusion chromatography (SEC), cation-exchange chromatography (CEC) and reversed-phase chromatography (RPC). The chapter summarizes the main features of ProDigest-LC, outlines the concepts which led to its development and demonstrates some of its prediction and simulation capabilities. Development of ProDigest-LC required accurate methods of retention time prediction in SEC, CEC, and RPC of peptides.
