Hi there,
I want to remove any ions (mostly Ca2+, amino acids) from my soluble protein extract preparation. I came across with idea of using C18 column which only binds to peptides and rest will pass through.
Can any body having experience with these columns confirm this. If this is not the case, what would they suggest for my experiment.
I appreciate any suggestion from my fellow researchers..
Cheers,
Raise
So long as the compounds you want to remove have no retention on C18, that is a good way of removing such impurities. The calcium ions can be removed in this way. You may need to use a gradient to resolve some compounds that have some retention on C18. I prefer an AQ type column, as it resists phase collapse. The link shows a compound being desalted which elutes at ~20% methanol from C18AQ. I've also used the technique to recover an NMR sample from DMSO-D6.
https://www.teledyneisco.com/en-us/liquidChromatography/Chromatography%20Documents/Application%20Notes/Desalting%20Samples%20with%20RediSep%20Rf%20Gold%20C18Aq%20Columns%20App%20Note.pdf
SEC desalting columns (separation based on molecular size) should work well for this purpose. C18 columns will bind proteins so you need to find a suitable solvent system for elution. Also you can precipitate proteins by an solvent to remove other soluble small molecules.
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