Extraction and stability of bovine serum albumin (BSA) using cholinium-based Good's buffers ionic liquids

• Novel ABS composed of GB-ILs/GB's/sucrose and PPG400 were investigated.
• The partition of a model protein, BSA, in these ABS was evaluated.
• BSA preferentially migrates to the GB-IL/GB/sucrose-rich phase.
• Extraction efficiencies of 100% of BSA were achieved in a single-step.
• The GB-ILs enhance the BSA stability.

Good's buffers ionic liquids (GB-ILs), composed of cholinium-based cations and Good's buffers anions, display self-buffering characteristics in the biological pH range, and their polarity and hydrophobicity can be easily tuned by a proper manipulation of their ions chemical structures. In this work, the extraction ability for bovine serum albumin (BSA) of aqueous biphasic systems (ABS) formed by polypropylene glycol 400 (PPG 400) and several GB-ILs was evaluated. ABS formed by PPG 400 and cholinium chloride ([Ch]Cl), GBs, and sucrose were also investigated for comparison purposes. It is shown that BSA preferentially migrates for the GB-IL-rich phase, with extraction efficiencies of 100%, achieved in a single-step. Dynamic light scattering, and circular dichroism (CD) and Fourier transform infrared (FTIR) spectroscopies were employed to evaluate the effect of the investigated cholinium-based GB-ILs on the BSA stability, and compared with results obtained for the respective GBs precursors, [Ch]Cl and sucrose, a well-known protein stabilizer. Molecular docking studies were also carried out to investigate on the binding sites of GB-IL ions to BSA. The experimental results confirm that BSA has a higher stability in GB-ILs than in any of the other compounds investigated.

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