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Flash Purification of Peptides - Part 2

19 January 2017

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. Depending on the quality of peptides synthesized, flash chromatography can be used for purification of peptides either as:

 

  1. the sole purification method (read more in AN112) or
  2. as a front end clean-up (AN113) prior to a final reversed phase HPLC polishing step

This application note (AN113) describes a variety of examples demonstrating how flash chromatography can be used as a front end clean-up method for crude synthetic peptides which can make the subsequent RP-HPLC step easier.

Conclusion

We have demonstrated that flash chromatography is a powerful technique to increase the purity of crude peptide samples. It can be used in tandem with RP-HPLC to quickly clean up peptide samples to limit the time performing the final purification step which expedites the peptide synthesis workflow from crude synthetic peptide to the final purified product.

The increased loading capacity of flash cartridges compared to prep RP-HPLC allows more peptide to be processed in a single injection and significant amounts of impurities can be removed that would require more time and effort to purify by RP-HPLC alone.

The Biotage® SNAP Bio 300 Å flash cartridges provided increased resolution with better separation and better recovery when purifying crude peptide mixtures in comparison to standard C18 flash cartridges. This enables faster and more efficient purification of peptides and larger molecules.

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