Application of reversed-phase liquid chromatography in separation and analysis of proteins and peptides

Reversed-phase liquid chromatography (RPLC) is based on the hydrophobic interaction, and has found increasing use in analysis and separation of bio-molecules due to its high resolution, reproducibility and recovery. In this review, the mechanism of RPLC is discussed and the applications of RPLC in the study of proteins and peptides are summarized. Adsorption and desorption of proteins by RPLC are described with an "on-off" mechanism, which states the interactions between the proteins, the molecules in the mobile phase and the stationary phase. The applications of RPLC are advanced in recent years, including protein and peptide purification, peptide mapping analysis, enzyme activity assay, conformational change detection and hydrophobic interaction study. The prospect of reversed-phase liquid chromatography in this field is anticipated.