Tandem electric field gradient focusing system for isolation and concentration of target proteins.
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.
Lin
Shu-Ling
SL
Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT 84602, USA.
Li
Yuanyuan
Y
Tolley
H Dennis
HD
Humble
Paul H
PH
Lee
Milton L
ML
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*
D011506:Proteins; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate