pH-regulated electroretention chromatography: towards a new method for the separation of proteins according to their isoelectric points.
The pH-dependent electroretention behavior of model proteins cytochrome c and ribonuclease A was studied in a hollow fiber arrangement, similar to that used in electrical field-flow fractionation. Field-induced immobilization of the proteins at the inner wall of the fiber was a function of the pH adjusted in the solution surrounding it, indicating that the pH inside the fiber lumen, relevant for protein migration, quickly equilibrates to the regulated value outside. A complete separation of the model proteins was achieved. Advantages of the principle as well as prospects for the development of a technique separating more than two protein species according to their isoelectric points are discussed.
Korlach
J
J
Mathematisch-Naturwissenschaftliche Fakultät I, Humboldt-Universität zu Berlin, Germany.
Hagedorn
R
R
Fuhr
G
G
D000818:Animals; D002417:Cattle; D005591:Chemical Fractionation / Q000379:methods; D002845:Chromatography / Q000295:instrumentation / Q000379:methods*; D003574:Cytochrome c Group / Q000302:isolation & purification; D019075:Electrophoresis, Capillary / Q000379:methods; D006736:Horses; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing / Q000379:methods; D007526:Isoelectric Point; D011506:Proteins / Q000302:isolation & purification*; D012259:Ribonuclease, Pancreatic / Q000302:isolation & purification
D003574:Cytochrome c Group; D011506:Proteins; D012259:Ribonuclease, Pancreatic