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      Increasing sample load for peptide purification with a step gradient

      Jun 10, 2021 2:32:20 PM / by Elizabeth Denton posted in Troubleshooting and Optimization, Scale-Up, Peptide Purification, flash chromatography


      We’ve all done it. Loaded a very small amount of our peptide onto an analytical, maybe semi-prep column, to see where the desired peak elutes, then create a shortened version of that gradient for the actual purification runs. A scouting run like the above mentioned gives the chemist a lot of information about the sample – resolution from impurities, potential gradient slopes to improve the separation, even loading capacity can be calculated.

      But how many times have you used a scouting run like this to figure out the purification gradient, only to have everything change when you increased your sample load? In today’s post, I highlight how easy scaling - aka increasing your sample load - can be accomplished when using reversed phase flash chromatography for peptide purification and will include some tips for success along the way.

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