The purification step is one of the main bottlenecks in the peptide synthesis workflow. Preparative RP-HPLC is normally the method of choice 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. Although there are a number of examples in the literature, flash chromatography is almost never considered as a technique for purification of synthetic peptides, as it is not perceived to be suitable for this application. However, with recent advances in flash using 20–25 micron spherical particles, making flash chromatography an efficient technique for synthetic peptide purification.
Biotage® SNAP Bio flash cartridges were developed with a small particle size (20 μm) and large pore size (300 Å) to provide increased resolution and effective separation of complex peptide mixtures. Flash chromatography can be used either as the sole purification method or as a front end clean-up prior to a final RP-HPLC step.
In part 1 of this application note we show a variety of examples demonstrating how flash chromatography can be used as the sole purification method for purifying crude synthetic peptides.
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