Peptide Mapping by Reverse-Phase High Pressure Liquid Chromatography

Reverse-phase high pressure liquid chromatography (HPLC) has proved to be an extremely versatile technique for rapid separation of peptides (reviewed in ref. 1). One of its uses is for peptide mapping, or "fingerprinting" (2-5) as an alternative procedure to the conventional two-dimensional separations on paper or thin-layer supports (See Chapter 21 ). Although the map obtained is one dimensional, the excellent resolving power of reverse-phase HPLC enables separation of the majority of peptides within a mixture. HPLC offers the advantages of high reproducibility, easy quantitation, and rapid analysis time, and is also suitable for automation.