A slow gradient approach for the purification of synthetic polypeptides by reversed phase high performance liquid chromatography

J Pept Sci. 2012 Sep;18(9):549-55. doi: 10.1002/psc.2432. Epub 2012 Jul 25.


Unquestionably, the purification of polypeptides by chromatographic methods is a considerable bottleneck in their preparation. Peptides synthesised by solid phase synthesis typically contain chromatographically similar impurities that complicate purification by reversed phase high performance liquid chromatography (HPLC) techniques. We report on the application of a slow gradient HPLC protocol that allows, in a single chromatographic step, the purification of hundreds of milligrammes of material. This technique was applied to an extensive collection of synthetic polypeptides some incorporating non-proteinogenic functionality. In all cases examined, the peptides were not only obtained in high purity peptides but were also recovered in multi-milligramme amounts.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatography, Reverse-Phase / methods*
  • Peptides / isolation & purification*


  • Peptides