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Whenever I synthesized a new peptide, I always ran a “scout run” – a small scale injection, usually with an analytical HPLC column – to both get an idea of the crude purity and also to identify a shorter, more optimal gradient for the actual purification. This strategy is still probably fine when you want to use reversed phase flash chromatography for your purification strategy, but is there a better way?
In today’s post, I’ll discuss using a scouting column to screen gradient conditions prior to peptide purification with reversed phase flash chromatography.
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Flash chromatography can be a challenging technique for peptide purification due to the lower resolution achieved with large particles. While some may see this as a disadvantage, the significantly greater loading capacity gives me reason to make this work. So how can I achieve the high purity levels often accessed using traditional reversed-phase HPLC methods?
In this post, I’ll discuss using a step gradient for peptide purification. Step gradients are commonly used in normal-phase small molecule purification and typically improve the purification efficiency while reducing the overall purification time. Continue reading How to purify peptides using a step gradient in flash column chromatography