Effects of ions on partitioning of serum albumin and lysozyme in aqueous two-phase systems containing ethylene oxide/propylene oxide co-polymers.
Aqueous two-phase systems composed of ethylene oxide/propylene oxide random co-polymers, EO30/PO70 or Ucon (EO50/PO50), in the top phase and dextran T500 in the bottom phase, have been studied. The cloud point diagram for EO30/PO70 in water solution was determined. EO30/PO70 has a cloud point of 32 degrees C at a concentration of 10% (w/w). The phase diagram for the system EO30/PO70-dextran T500-water was determined. Salt effects have been studied on the partitioning of two model proteins, bovine serum albumin and hen egg white lysozyme, in EO30/PO70-dextran and Ucon-dextran systems. Ions with different hydrophobicity, i.e., with different position in the Hofmeister or lyotropic series, were investigated with reference to their effect on protein partition. The counterion hydrophobicity was shown to have a strong influence on the partitioning of BSA and lysozyme. Most extreme partitioning was obtained with hydrophobic (chaotropic) ions like CIO4- and I-. A comparison of protein partitioning between PEG-dextran and EO30/PO70-dextran has been done. A more extreme protein partitioning was obtained in the EO30/PO70-dextran containing system. Temperature-induced phase separation was studied with EO30/PO70 at 45 degrees C. Both BSA and lysozyme were completely partitioned to the water phase formed above the cloud point of EO30/PO70. Model calculations, based on Flory-Huggins theory of polymer solutions, have been done which could reproduce the salt effect on the protein partitioning in aqueous-two phase system.
Johansson
H O
HO
Department of Biochemistry, Lund University, Sweden.
Lundh
G
G
Karlström
G
G
Tjerneld
F
F
D001965:Bromides / Q000737:chemistry; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D005998:Glycine / Q000737:chemistry; D007477:Ions; D008956:Models, Chemical; D009113:Muramidase / Q000737:chemistry*; D010472:Perchlorates / Q000737:chemistry; D011095:Polyethylenes / Q000737:chemistry*; D011126:Polypropylenes / Q000737:chemistry*; D012710:Serum Albumin, Bovine / Q000737:chemistry*; D012965:Sodium Chloride / Q000737:chemistry; D017670:Sodium Compounds / Q000737:chemistry; D012974:Sodium Iodide / Q000737:chemistry; D014867:Water / Q000737:chemistry*
D001965:Bromides; D007477:Ions; D010472:Perchlorates; D011095:Polyethylenes; D011126:Polypropylenes; D017670:Sodium Compounds; D014867:Water; D012710:Serum Albumin, Bovine; D012965:Sodium Chloride; C078661:UCON 50-HB-5100; C031068:sodium perchlorate; D009113:Muramidase; D012974:Sodium Iodide; C027938:sodium bromide; D005998:Glycine