Synthetic peptides are typically purified by reversed-phase HPLC (RP-HPLC) chromatography using a mobile phase containing an acidic modifier.  Although capable of very high resolution, RP-HPLC is often limited by column loading capacity and therefore demanding a significant time investment for peptide purification. As an alternatively strategy, reversed-phase flash column chromatography can also be used to purify synthetic peptides.  The larger particle size used in flash column chromatography enables much large loading capacity, thereby significantly reducing the time required for peptide purification, albeit at a lower resolution. <br/><br/>Herein, we report a strategy to rapidly purify peptides to a high degree of purity using reversed-phase flash column chromatography.  We demonstrate that increasing the mobile phase pH for a mixed solution of linear and disulfide-mediated cyclic oxytocin, a solution not resolvable under standard reversed phase conditions, enables separation and purification of the desired cyclic peptide.  This strategy is then applied to the purification of GLP-1, a 37 amino acid peptide demanding exceptionally high purity for use in biological studies.  These results clearly demonstrate that increasing the mobile phase pH can enable rapid peptide purification, yielding a highly pure product.

Improving peptide purification via flash column chromatography by modulating mobile phase pH

253rd National Meeting (2017)

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