Study of the applicability of non-conventional aqueous two-phase systems in counter-current and centrifugal partition chromatography.
Aqueous two-phase systems composed of imidazolium-based ionic liquids and phosphate salts were evaluated for their applicability in liquid-liquid chromatography. The influence of the nature of ionic liquid anion and cation on the partitioning of bovine serum albumin, lysozyme and myoglobin was investigated. A mixture of K2HPO4 and KH2PO4 in a ratio of 1.82:1 wt/wt was used in all of the tested biphasic systems to adjust the pH to a range of 7-8. The results show that more hydrophobic cations decrease the partition coefficients of the proteins in the biphasic systems and outweigh the effect of the anion on the distribution of the macromolecules. Viscosities and densities of the biphasic systems were in a suitable range for liquid-liquid chromatography. Even though the partition coefficients were too high for a conventional batch operation mode, these aqueous two-phase systems show favorable properties for protein capturing in liquid-liquid chromatographic columns. Additionally, the possible application of ionic liquids as modifiers in polyethylene glycol (PEG)-based aqueous two-phase systems was investigated. It could be demonstrated that ionic liquids alter the partition coefficients of the proteins.
Biothermodynamics, TUM School of Life Sciences Weihenstephan, Technische Universität München, D-85354 Freising, Germany.
Franziska
F
Bezold
Biothermodynamics, TUM School of Life Sciences Weihenstephan, Technische Universität München, D-85354 Freising, Germany.
Johannes
J
Goll
Biothermodynamics, TUM School of Life Sciences Weihenstephan, Technische Universität München, D-85354 Freising, Germany. Electronic address: mirjana.minceva@tum.de.
Mirjana
M
Minceva
D000818:Animals; D002417:Cattle; D002498:Centrifugation; D003377:Countercurrent Distribution / Q000379:methods*; D057927:Hydrophobic and Hydrophilic Interactions; D007093:Imidazoles / Q000737:chemistry; D052578:Ionic Liquids / Q000737:chemistry*; D009113:Muramidase / Q000737:chemistry / Q000302:isolation & purification*; D009211:Myoglobin / Q000737:chemistry / Q000302:isolation & purification*; D011092:Polyethylene Glycols / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000302:isolation & purification*; D014783:Viscosity; D014867:Water / Q000737:chemistry*
D007093:Imidazoles; D052578:Ionic Liquids; D009211:Myoglobin; D014867:Water; D012710:Serum Albumin, Bovine; D011092:Polyethylene Glycols; C029899:imidazole; D009113:Muramidase
Aqueous two-phase system; Centrifugal partition chromatography; Counter-current chromatography; Ionic liquid; Protein separation