Ion Exchange Chromatography - How to Choose the IEX Buffer?
The buffer ion should have the same charge as the functional groups on the IEX medium. For anion exchange, choose a buffer with a pH 0.5-1 units above the isoelectric point (pI) because as the pI increases, the protein binds more tightly. For cation exchange, choose a buffer with a pH 0.5-1 units below the pI because as the pI decreases, the protein binds more tightly. If both anion and cation exchange are conducted simultaneously, the stability of the protein needs to be considered, as many proteins are unstable when the pH is close to their isoelectric point (pI).
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