title: Tandem electric field gradient focusing system for isolation and concentration of target proteins.
abstract: Two electric field gradient focusing (EFGF) systems, one based on a hollow dialysis fiber and the other based on a shaped ionically conductive polymer were serially integrated to trap and concentrate selected proteins while simultaneously desalting and removing other unwanted proteins from the sample. A series of experiments were performed to test the EFGF systems individually and after integration. Online concentration of amyloglucosidase indicated a concentration limit of detection of approximately 20 ng mL(-1) (200 pM) from a sample volume of 100 microL. Concentration of human alpha1-acid glycoprotein with simultaneous removal of human serum albumin was also demonstrated. Elimination of small buffer components while concentrating trypsin inhibitor, and selective concentration and separation of myoglobin from a mixture were performed using the integrated EFGF system.
authors.affiliation: Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT 84602, USA.
authors.forename: Shu-Ling
authors.initials: SL
authors.lastname: Lin
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Li
authors.forename: H Dennis
authors.initials: HD
authors.lastname: Tolley
authors.forename: Paul H
authors.initials: PH
authors.lastname: Humble
authors.forename: Milton L
authors.initials: ML
authors.lastname: Lee
mesh_terms: D000818:Animals; D002417:Cattle; D002645:Chickens; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry; D007525:Isoelectric Focusing / Q000379:methods*; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D011506:Proteins; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate