Peptide mapping by reversed phase chromatography

Reversed phase chromatography is widely used in analytical biotechnology and biochemistry for the separation of fragments obtained by the proteolytic digest of a protein. The peptide maps, thus generated, are used to assess the integrity of amino acid sequences of recombinant DNA proteins and to gain information on the primary sequence of newly discovered proteins. This article briefly reviews this analytical procedure and discusses some aspects related to the contribution of column packing material and mobile phase composition on the separation of proteolytic peptides by reversed phase chromatography.