High-performance hydrophobic interaction chromatography of proteins on reversed-phase supports coated with non-ionic surfactants of polyoxyethylene type. Purification of a fungal aspartic proteinase.
On coating reversed-phase supports with polyoxyethylene-type non-ionic surfactants, proteins are no longer retained on such supports at moderate or low ionic strength, but they are retained at high ionic strength and can be desorbed by a decreasing ionic strength gradient. These reversibly modified supports were used for hydrophobic interaction chromatography (HIC). The proteins probably interact with the polyoxyethylene tail of the non-ionic surfactant while the hydrophobic part of the surfactant anchors the surfactant to the reversed-phase support by interactions with its alkane coverage. Although the interactions between non-ionic surfactant and reversed-phase support are non-covalent and the HIC mobile phases contained no surfactant, the modified columns were stable and could be used repeatedly. A surfactant-modified reversed-phase column provided a rapid, efficient, one-step purification of a fungal aspartic proteinase from a commercial crude preparation.
Sing
Y L
YL
INSERM U 160, Hospital Beaujon, Clichy, France.
Kroviarski
Y
Y
Cochet
S
S
Dhermy
D
D
Bertrand
O
O
D016282:Aspartic Acid Endopeptidases / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D011092:Polyethylene Glycols; D012233:Rhizopus / Q000201:enzymology*; D013501:Surface-Active Agents
D013501:Surface-Active Agents; D011092:Polyethylene Glycols; D016282:Aspartic Acid Endopeptidases