Partitioning in Aqueous Two-Phase Systems: Fundamentals, Applications and Trends

Over the last years, aqueous two-phase systems (ATPS) regained an increasing interest due to their potential in the downstream processing of biomolecules. After many years with only a few articles published, a lot of effort and work has been put into studying these systems for the partitioning of a range of compounds including proteins, organic low-molecular weight molecules or metal ions. Although several research and review articles appeared, a background review on ATPS partitioning fundamentals is needed. In this article, partitioning theories and main effects of several important factors for partitioning, such as molecular weight of the polymer, effect of added salts, pH, electrical charges, and temperature on phase diagrams, tie-line lengths, interfacial tension and settling time of the two aqueous phases are extensively reviewed. The trend in ATPS research is given compiling the recent 2008–2013 research articles published in the field.

[1]  Marco Rito-Palomares,et al.  Continuous aqueous two-phase systems devices for the recovery of biological products , 2014 .

[2]  T. A. Hatton,et al.  Protein partitioning in two-phase aqueous polymer systems. , 1989, Biotechnology and bioengineering.

[3]  S. Mustafa,et al.  Optimization of Serine Protease Purification from Mango (Mangifera indica cv. Chokanan) Peel in Polyethylene Glycol/Dextran Aqueous Two Phase System , 2012, International journal of molecular sciences.

[4]  Yanglong Zhang,et al.  Partitioning of biomolecules in aqueous two-phase systems of polyethylene glycol and nonionic surfactant , 2012 .

[5]  F. Tjerneld,et al.  Effect of electrolytes and surfactants on the thermoseparation of an ethylene oxide-propylene oxide random copolymer in aqueous solution. , 1998, Journal of chromatography. B, Biomedical sciences and applications.

[6]  Ana M Azevedo,et al.  Chromatography-free recovery of biopharmaceuticals through aqueous two-phase processing. , 2009, Trends in biotechnology.

[7]  Yang Liu,et al.  Phase equilibrium and protein partitioning in aqueous micellar two-phase system composed of surfactant and polymer , 2012 .

[8]  Ana M. Azevedo,et al.  Partitioning of human antibodies in polyethylene glycol–sodium citrate aqueous two-phase systems , 2009 .

[9]  Folke Tjerneld,et al.  [1] Phase diagrams , 1994 .

[10]  A. Salabat,et al.  Amino acids partitioning in aqueous two-phase system of polypropylene glycol and magnesium sulfate. , 2007, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[11]  M. Aires-Barros,et al.  Affinity partitioning of human antibodies in aqueous two-phase systems. , 2007, Journal of chromatography. A.

[12]  M. Rito‐Palomares,et al.  Aqueous two-phase systems strategies to establish novel bioprocesses for stem cells recovery , 2014, Critical reviews in biotechnology.

[13]  Zhiyong Li,et al.  Partitioning behavior of amino acids in aqueous two-phase systems formed by imidazolium ionic liquid and dipotassium hydrogen phosphate. , 2012, Journal of chromatography. A.

[14]  M. Aires-Barros,et al.  Purification of human immunoglobulin G by thermoseparating aqueous two-phase systems. , 2008, Journal of chromatography. A.

[15]  Marco Rito-Palomares,et al.  Aqueous two-phase affinity partitioning systems: current applications and trends. , 2012, Journal of chromatography. A.

[16]  T. A. Hatton,et al.  Thermodynamics of the partitioning of biomaterials in two-phase aqueous polymer systems: comparison of lattice model to experimental data , 1989 .

[17]  T. Ling,et al.  Recovery of glyceraldehyde 3-phosphate dehydrogenase from an unclarified disrupted yeast using aqueous two-phase systems facilitated by distribution analysis of radiolabelled analytes , 2012 .

[18]  Two new crystal forms of the choline-binding domain of the major pneumococcal autolysin: insights into the dynamics of the active homodimer. , 2002, Journal of molecular biology.

[19]  P. Albertsson.  1 – History of Aqueous Polymer Two-Phase Partition , 1985 .

[20]  P. Albertsson,et al.  Partition of Cell Particles and Macromolecules , 1986 .

[21]  M. Aires-Barros,et al.  Downstream processing of human antibodies integrating an extraction capture step and cation exchange chromatography. , 2009, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[22]  J. Teixeira,et al.  Enzyme purification with aqueous two-phase systems: comparison between systems composed of pure polymers and systems composed of crude polymers. , 1996, Journal of chromatography. B, Biomedical applications.

[23]  Juan A. Asenjo,et al.  Theoretical and Experimental Evaluation of Hydrophobicity of Proteins to Predict their Partitioning Behavior in Aqueous Two Phase Systems: A Review , 2010 .

[24]  D. Wei,et al.  Partitioning behaviour of cephalexin and 7‐aminodeacetoxicephalosporanic acid in PEG/ammonium sulfate aqueous two‐phase systems , 2001 .

[25]  J. Coutinho,et al.  Characterization of aqueous biphasic systems composed of ionic liquids and a citrate-based biodegradable salt , 2012 .

[26]  J. Prausnitz,et al.  Molecular thermodynamics of aqueous two-phase systems for bioseparations , 1988 .

[27]  B. Zaslavsky,et al.  Solvent properties governing protein partitioning in polymer/polymer aqueous two-phase systems. , 2011, Journal of chromatography. A.

[28]  D. Lin,et al.  Thermodynamics of aqueous two-phase systems—the effect of polymer molecular weight on liquid–liquid equilibrium phase diagrams by the modified NRTL model , 1998 .

[29]  J. Asenjo,et al.  Partitioning and purification of monoclonal antibodies in aqueous two-phase systems. , 1996, Bioseparation.

[30]  M. Mokhtar,et al.  Recovery of Bacillus cereus cyclodextrin glycosyltransferase and recycling of phase components in an aqueous two-phase system using thermo-separating polymer , 2012 .

[31]  D. Prazeres,et al.  Purification of plasmid DNA with aqueous two phase systems of PEG 600 and sodium citrate/ammonium sulfate , 2009 .

[32]  X. Joulia,et al.  Liquid-Liquid Equilibria of the Ternary Systems 3-Methyl-1-butanol + Ethanol + Water and 2-Methyl-1-propanol + Ethanol + Water at 293.15 K , 2008 .

[33]  V. Vasić,et al.  Evaluation of possibility of textile dye removal from wastewater by aqueous two-phase extraction , 2013 .

[34]  T. Ling,et al.  Selective partition of plasmid DNA and RNA in aqueous two-phase systems by the addition of neutral salt , 2009 .

[35]  Marco Rito-Palomares,et al.  Rotavirus-like particles primary recovery from insect cells in aqueous two-phase systems. , 2006, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[36]  M. Rito‐Palomares,et al.  Kinetics of phase separation under different process and design parameters in aqueous two-phase systems. , 2000, Journal of chromatography. B, Biomedical sciences and applications.

[37]  H Cabezas,et al.  Theory of phase formation in aqueous two-phase systems. , 1996, Journal of chromatography. B, Biomedical applications.

[38]  J. Persson,et al.  Aqueous two-phase systems with smart polymers , 2002 .

[39]  Jyh-Ping Chen,et al.  Lactose hydrolysis by β-galactosidase in aqueous two-phase systems , 1991 .

[40]  D. Peričin,et al.  THE EFFECT OF SULPHATES ON PARTITIONING OF PECTINASES IN AQUEOUS TWO-PHASE SYSTEMS , 2004 .

[41]  A. J. Meirelles,et al.  Phase equilibrium and protein partitioning in aqueous mixtures of maltodextrin with polypropylene glycol , 2001 .

[42]  Selvaraj Raja,et al.  Aqueous Two Phase Systems for the Recovery of Biomolecules –A Review , 2012 .

[43]  J. Hsu,et al.  Correlation of protein partitioning in aqueous polymer two-phase systems , 1990 .

[44]  M. C. Madhusudhan,et al.  Aqueous two phase extraction of invertase from baker's yeast: Effect of process parameters on partitioning , 2011 .

[45]  K. Ratanapongleka.  Recovery of Biological Products in Aqueous Two Phase Systems , 2010 .

[46]  G. Zanin,et al.  Partitioning of Porcine Pancreatic Lipase in a Two-Phase Systems of Polyethylene Glycol/Potassium Phosphate Aqueous , 2010, Applied biochemistry and biotechnology.

[47]  E. Tambourgi,et al.  New aqueous two-phase system based on cashew-nut tree gum and poly(ethylene glycol). , 2000, Journal of chromatography. B, Biomedical sciences and applications.

[48]  G. Picó,et al.  Partitioning features of bovine trypsin and alpha-chymotrypsin in polyethyleneglycol-sodium citrate aqueous two-phase systems. , 2007, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[49]  C. Glatz,et al.  Aqueous two-phase extraction for protein recovery from corn extracts. , 2007, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[50]  Tingyue Gu,et al.  Liquid-liquid partitioning methods for bioseparations , 2000 .

[51]  A. Alfani,et al.  Phase diagram, rheology and interfacial tension of aqueous mixtures of Na-caseinate and Na-alginate , 2004 .

[52]  G. Johansson.  [3] Partitioning procedures and techniques: Small molecules and macromolecules , 1994 .

[53]  L. Mei,et al.  Densities and Viscosities of Polyethylene Glycol + Salt + Water Systems at 20 .degree.C , 1995 .

[54]  T. Panda,et al.  Aqueous two-phase: the system of choice for extractive fermentation , 2000, Applied Microbiology and Biotechnology.

[55]  Murni M. Ahmad,et al.  Towards optimal aqueous two‐phase extraction system flowsheets for protein purification , 2013 .

[57]  Analysis of amino acid-water interactions by partitioning in aqueous two-phase systems. I--amino acids with non-polar side-chains. , 2013, Journal of chromatography. A.

[58]  Yongsheng Yan,et al.  Liquid–liquid equilibrium composed of imidazolium tetrafluoroborate ionic liquids + sodium carbonate aqueous two-phase systems and correlation at (288.15, 298.15, and 308.15) K , 2011 .

[59]  R. Guzman,et al.  Affinity Partitioning of Metal Ions in Aqueous Biphasic Systems: Experimental and Theoretical Aspects , 1995 .

[60]  M. Aires-Barros,et al.  Integrated process for the purification of antibodies combining aqueous two-phase extraction, hydrophobic interaction chromatography and size-exclusion chromatography. , 2008, Journal of chromatography. A.

[61]  M. Rodrigues,et al.  Evaluation of the effects of the parameters involved in the purification of clavulanic acid from fermentation broth by aqueous two-phase systems , 2009, Bioprocess and biosystems engineering.

[62]  L. D. Silva,et al.  Surface Excess Enthalpy of PEO + Salt +Water and L35 + Salt + Water Aqueous Two-Phase Systems , 2009 .

[63]  Zhiyong Li,et al.  Partitioning Behavior of Wastewater Proteins in Some Ionic Liquids-Based Aqueous Two-Phase Systems , 2012 .

[64]  A. Santhiagu,et al.  Technological aspects of extractive fermentation using aqueous two-phase systems , 2003 .

[65]  T. A. Hatton,et al.  Thermodynamics of the separation of biomaterials in two-phase aqueous polymer systems: effect of the phase-forming polymers , 1987 .

[66]  F. Leermakers,et al.  Comment on: Thermodynamics of the separation of biomaterials in two-phase aqueous polymer systems: Effect of the phase-forming polymers. , 1988 .

[67]  L. A. van der Wielen,et al.  Poly(ethylene glycol)-salt aqueous two-phase systems with easily recyclable volatile salts. , 1998, Journal of chromatography. B, Biomedical sciences and applications.

[68]  Marco Rito-Palomares,et al.  Low-abundant protein extraction from complex protein sample using a novel continuous aqueous two-phase systems device. , 2013, Journal of separation science.

[69]  A. Salabat.  The influence of salts on the phase composition in aqueous two-phase systems: experiments and predictions , 2001 .

[70]  S. Sandler,et al.  Aqueous Two-Phase Systems. 2. Protein Partitioning , 1994 .

[71]  F. Tjerneld,et al.  Temperature-induced phase partitioning of peptides in water solutions of ethylene oxide and propylene oxide random copolymers. , 1997, Biochimica et biophysica acta.

[72]  R. Dinarvand,et al.  Evaluation of recombinant phenylalanine dehydrogenase behavior in aqueous two-phase partitioning , 2007 .

[73]  G. Picó,et al.  Extraction of trypsin from bovine pancreas by applying polyethyleneglycol/sodium citrate aqueous two-phase systems. , 2009, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[74]  B. Cavada,et al.  Liquid-Liquid Equilibrium Data for Aqueous Two-Phase Systems Composed of Ethylene Oxide Propylene Oxide Copolymers , 2011 .

[75]  B. Zaslavsky,et al.  Solvent properties governing solute partitioning in polymer/polymer aqueous two-phase systems: nonionic compounds. , 2010, The journal of physical chemistry. B.

[76]  P. Bahadur,et al.  Effect of different additives on the cloud point of a polyethylene oxide-polypropylene oxide-polyethylene oxide block copolymer in aqueous solution , 2002 .

[77]  A. Pacek,et al.  The effect of temperature and composition on the interfacial tension and rheology of separated phases in gelatin/pullulan mixtures , 2005 .

[78]  P. Albertsson.  Aqueous biphasic systems. Properties and applications in bioseparation.. , 1995 .

[79]  Ann Schluederberg,et al.  Partition of Cell Particles and Macromolecules , 1973, The Yale Journal of Biology and Medicine.

[80]  Carol K. Hall,et al.  Interfacial tension of polyethyleneglycol-dextran-water systems: influence of temperature and polymer molecular weight , 1990 .

[81]  J. Coutinho,et al.  Extraction of tetracycline from fermentation broth using aqueous two-phase systems composed of polyethylene glycol and cholinium-based salts , 2013 .

[82]  B. Nerli,et al.  Molecular features determining different partitioning patterns of papain and bromelain in aqueous two-phase systems. , 2013, International journal of biological macromolecules.

[83]  P. A. P. Filho,et al.  Experimental determination and thermodynamic modeling of phase equilibrium and protein partitioning in aqueous two-phase systems containing biodegradable salts , 2013 .

[84]  J. Cabral,et al.  Cell partitioning in aqueous two-phase polymer systems. , 2007, Advances in biochemical engineering/biotechnology.

[85]  F. Deive,et al.  Sodium carbonate as phase promoter in aqueous solutions of imidazolium and pyridinium ionic liquids , 2011 .

[86]  Yongsheng Yan,et al.  Liquid−Liquid Equilibrium of Potassium Phosphate/Potassium Citrate/Sodium Citrate + Ethanol Aqueous Two-Phase Systems at (298.15 and 313.15) K and Correlation , 2010 .

[87]  D. Forciniti,et al.  Effect of glycosylation on the partition behavior of a human antibody in aqueous two‐phase systems , 2013, Biotechnology progress.

[88]  Ana M Azevedo,et al.  Continuous purification of antibodies from cell culture supernatant with aqueous two-phase systems: from concept to process. , 2013, Biotechnology journal.

[89]  T. Franco,et al.  Extraction in aqueous two-phase systems of alkaline xylanase produced by Bacillus pumilus and its application in kraft pulp bleaching. , 2000, Journal of chromatography. B, Biomedical sciences and applications.

[90]  G. Johansson.  Comparison of two aqueous biphasic systems used for the partition of biological material. , 1978, Journal of chromatography.

[91]  Ana M Azevedo,et al.  Optimisation of aqueous two-phase extraction of human antibodies. , 2007, Journal of biotechnology.

[92]  P. D. Belur,et al.  Development and Evaluation of PEG-Lithium Citrate Salt Based Aqueous Two Phase System and Its Application in Partitioning of Proteins from Fish Industry Effluent , 2012 .

[93]  A. Pacek,et al.  Interfacial tension in phase-separated gelatin/dextran aqueous mixtures. , 2002, Journal of colloid and interface science.

[94]  B. Zaslavsky,et al.  Salt effects on solvent features of coexisting phases in aqueous polymer/polymer two-phase systems. , 2012, Journal of chromatography. A.

[95]  C. D. de Gooijer,et al.  Hybridoma and CHO Cell Partitioning in Aqueous Two‐Phase Systems , 1996, Biotechnology progress.

[96]  G. Picó,et al.  Application of the aqueous two-phase systems of ethylene and propylene oxide copolymer-maltodextrin for protein purification. , 2005, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[97]  F. Tjerneld,et al.  Application of temperature-induced phase partitioning at ambient temperature for enzyme purification. , 1994, Journal of chromatography. A.

[98]  J A Asenjo,et al.  Phase separation rates of aqueous two‐phase systems: Correlation with system properties , 2002, Biotechnology and bioengineering.

[99]  E. Tambourgi,et al.  Partition of trypsin in aqueous two-phase systems of poly(ethylene glycol) and cashew-nut tree gum , 2002 .

[100]  Reinhard Lipowsky,et al.  Concentration dependence of the interfacial tension for aqueous two-phase polymer solutions of dextran and polyethylene glycol. , 2012, Langmuir : the ACS journal of surfaces and colloids.

[101]  A. Salabat,et al.  Partitioning of Amino Acids in Surfactant Based Aqueous Two-Phase Systems Containing the Nonionic Surfactant (Triton X-100) and Salts , 2011 .

[102]  T. Ramasami,et al.  Aqueous two-phase poly(ethylene glycol)–poly(acrylic acid) system for protein partitioning: Influence of molecular weight, pH and temperature , 2008 .

[103]  B. Mannervik,et al.  Effects of salts on the partition of proteins in aqueous polymeric biphasic systems. , 1974, Acta chemica Scandinavica. Series B: Organic chemistry and biochemistry.

[104]  M. Aires-Barros,et al.  Aqueous two-phase systems: A viable platform in the manufacturing of biopharmaceuticals. , 2010, Journal of chromatography. A.

[105]  Geoffrey V.F. Seaman,et al.  The effects of salts on the interfacial tension of aqueous dextran poly(ethylene glycol) phase systems , 1984 .