Despite the improvements in automated solid phase peptide synthesis (SPPS), such as the use of elevated temperatures enabling peptides of greater complexity to be synthesized routinely, the purification step is one of the main bottlenecks in the peptide synthesis workflow. Preparative reversed-phase HPLC is normally the method of choice for purification of synthetic peptides, but is limited by small loading amounts, long separation times, poor recoveries and high costs. In addition, crude synthetic peptides contain impurities with retention characteristics very similar to the target peptide which can present additional purification challenges.
Can you use anything else to achieve the necessary purity while maintaining a desirable yield and throughput? Here we demonstrate that high performance flash chromatography (HPFC) can be used as an efficient technique for the purification of synthetic peptides. We present data showing examples where high performance flash chromatography is used either as the sole method of peptide purification or as part of a multi-step process to initially remove most of the undesired impurities by HPFC prior to a final “polishing” step by RP-HPLC.
High performance flash chromatography allows more peptide to be processed in a single injection and significant amount of impurities can be removed that would require more time and effort to purify by RP-HPLC alone.