Protein separation and purification in neat dimethyl sulfoxide.
Pure DMSO (instead of water) is used as the reaction medium for protein separations. It is shown that common extracellular proteins (i) have high solubility in DMSO (1-50 mg/ml), (ii) do not irreversibly inactivate in this solvent, and (iii) can adsorb onto carboxymethyl cellulose in DMSO and be subsequently fully desorbed in this solvent by inorganic salts. Ion-exchange chromatography on this resin in DMSO has been used to purify bovine pancreatic trypsin and to separate it from hen egg-white lysozyme in their mixture. Another approach to protein separation in DMSO, fractional precipitation with ethyl acetate (which does not dissolve proteins), has been verified with a mixture of bovine pancreatic chymotrypsinogen and chicken egg ovalbumin.
Chang
N
N
Department of Chemistry, Massachusetts Institute of Technology, Cambridge 02139.
Hen
S J
SJ
Klibanov
A M
AM
D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange / Q000379:methods; D004121:Dimethyl Sulfoxide; D004798:Enzymes / Q000302:isolation & purification* / Q000378:metabolism; D007202:Indicators and Reagents; D007700:Kinetics; D009113:Muramidase / Q000302:isolation & purification; D011506:Proteins / Q000302:isolation & purification*; D012995:Solubility; D014357:Trypsin / Q000302:isolation & purification
D004798:Enzymes; D007202:Indicators and Reagents; D011506:Proteins; D009113:Muramidase; D014357:Trypsin; D004121:Dimethyl Sulfoxide