Choline-like ionic liquid-based aqueous two-phase extraction of selected proteins

Abstract

In this paper, an ionic liquid-based aqueous two-phase system (ATPs) has been developed for the effective extraction of bovine serum albumin (BSA). Compared with the effects of three different ionic liquids used in an aqueous two-phase system, a choline-like ionic liquid was selected as the suitable solution for bovine serum albumin. Extraction efficiencies of BSA were found to be dependant on the temperature, time, IL mass and the salt content. Based on single-factor experiments, these significant operating parameters of the ILs-ATP were respectively optimized. Experimental results show that 92.03–100.03% of the proteins could be extracted into the ionic liquid-rich phase in a single-step extraction. Repeatability, precision and stability experiments were investigated and the RSDs are lower than 2.3%, 1.7% and 4.7% respectively. Experiments indicated that intermolecular forces, the micelle phenomenon and the salting-out effect play important roles in the transfer process, and the aggregation phenomenon was the main driving force for the separation. Conformation of the protein was not affected after extraction into the upper ionic liquid-rich phase according to the determination by UV-visible (UV-vis), fluorescence spectra, dynamic light scattering (DLS), Fourier transform infrared (FT-IR) spectroscopy and TEM. It is suggested that the method of choline-like ILs-based ATPs for the selective separation of protein would have potential applications in bio-analysis and bio-separation.