title: Preparative-scale isoelectric trapping separations using a modified Gradiflow unit.
abstract: The Gradiflow BF200 preparative electrophoretic unit (Gradipore), which has been developed for size-based and charge-sign-based protein separations and in which the hydraulic flow path of the recirculating sample stream in the separation cartridge is orthogonal to the electric field, has been modified to carry out binary protein separations using the principles of isoelectric trapping. The disposable separation cartridge contained three isoelectric membranes which, along with the cartridge holder, formed the anode and cathode compartments and the anodic and cathodic separation compartments. The utility of the modified instrument was demonstrated by effecting a binary separation of chicken egg white across an isoelectric point 5.5 isoelectric membrane. The desalting and subsequent binary separation steps proved to be remarkably rapid, due to the favorable combination of short electrophoretic path, high electric field strength and large effective isoelectric membrane surface area.
authors.affiliation: Gradipore, French's Forest, NSW 2086, Australia.
authors.forename: David
authors.initials: D
authors.lastname: Ogle
authors.forename: Agnes
authors.initials: A
authors.lastname: Ho
authors.forename: Toby
authors.initials: T
authors.lastname: Gibson
authors.forename: Dennis
authors.initials: D
authors.lastname: Rylatt
authors.forename: Evan
authors.initials: E
authors.lastname: Shave
authors.forename: Peniel
authors.initials: P
authors.lastname: Lim
authors.forename: Gyula
authors.initials: G
authors.lastname: Vigh
mesh_terms: D007525:Isoelectric Focusing / Q000295:instrumentation / Q000379:methods*; D011506:Proteins / Q000302:isolation & purification
chemical_list: D011506:Proteins