Evaluation of ammonium acetate as a volatile buffer for high-performance hydrophobic-interaction chromatography

Abstract

Hydrophobic-interaction chromatography (HIC) is a widely used technique for the separation of proteins without denaturation. In HIC, although, ammonium sulphate or sodium sulphate buffer is generally used as an eluent, volatile buffers such as ammonium acetate and ammonium formate seem to be advantageous in order to simplify the subsequent procedures including desalting. Therefore, the applicability of ammonium acetate buffer was evaluated, as a representative of volatile buffers for HIC, with respect to effects on the retention and peak broadening of proteins. Several proteins were successfully separated under the optimized conditions using volatile ammonium acetate buffer.