What kind of buffer should I choose for ion exchange chromatography (IEX)?
The ions in the buffer should have the same charge as the functional groups on the IEX medium. For anion exchange, choose a buffer that is 0.5-1 pH units above the isoelectric point (pI) because proteins bind more tightly as the pI increases. For cation exchange, choose a buffer that is 0.5-1 pH units below the pI because proteins bind more tightly as the pI decreases. Whether it is anion or cation exchange, it is important to consider the protein's stability window, as many proteins become unstable when the pH is close to their isoelectric point (pI).
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