Influence of transport properties in electric field gradient focusing.
Miniaturized devices for electric field gradient focusing (EFGF) were developed that consist of a cylindrical separation channel surrounded by an acrylic-based polymer hydrogel. The ionic transport properties of the hydrogel enable the manipulation of the electric field inside the separation channel. A changing cross-section design was used in which the hydrogel is shaped such that an electric field gradient is established in the separation channel. One of the challenges with this type of EFGF device has been that experimental resolution between protein analytes is lower than theoretically predicted. In order to investigate this phenomenon, a mathematical transport model was developed using FEMLAB. Model results and experimental observations showed that the reduced performance was caused by concentration gradients formed in the EFGF channel, and that these concentration gradients were the result of an imbalance in cation transport between the open separation channel and the hydrogel. Removing acidic impurities from the monomers that form the hydrogel reduced this tendency and improved the resolution. These transport-induced concentration gradients can be used to establish electric field gradients that may be useful for sample pre-concentration. Both the results of simulation and experiments demonstrate how transport-induced concentration gradients lead to the establishment of electric field gradients.
Humble
Paul H
PH
Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT 84602, USA.
Harb
John N
JN
Tolley
H Dennis
HD
Woolley
Adam T
AT
Farnsworth
Paul B
PB
Lee
Milton L
ML
D004586:Electrophoresis / Q000379:methods*; D020342:Finite Element Analysis; D016650:Fluorescein-5-isothiocyanate; D049452:Green Fluorescent Proteins / Q000737:chemistry; D017136:Ion Transport; D008956:Models, Chemical; D010455:Peptides / Q000737:chemistry; D010799:Phycoerythrin / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry
D010455:Peptides; D011994:Recombinant Proteins; C485184:enhanced green fluorescent protein; D010799:Phycoerythrin; D049452:Green Fluorescent Proteins; D016650:Fluorescein-5-isothiocyanate