ABSTRACT
Traditionally, high-performance liquid chromatography (HPLC) of biopolymers is carried out by using stationary phases and chromatographic systems similar to those originally developed for the separation of small molecules. Thus, in HPLC of biological macromolecules, such as proteins and nucleic acids, microparticulate stationary phases made with macroporous supports are used and their separation is accomplished by reversed-phase, ion-exchange, or other types of chromatography under gradient elution conditions. Reversed-phase chromatography of peptides is a powerful technique for peptide mapping, for isolation of peptides, to determine amino acid sequence, for studies on microheterogeneity, mutational variants, and other isomers, as well as for study of glycosylation sites in proteins. Columns packed with a suitable micropellicular stationary phase are eminently suitable for rapid chromatographic separation of peptides, including peptide mapping. Biospecific pellicular sorbents appear to be ideal for analytical affinity separations, since they allow maximum exposure of the affinity ligate at the surface to interact with the molecules to be separated.
