Multistage Isoelectric Focusing in a Polymeric Microfluidic Chip
- 12 November 2005
- journal article
- research article
- Published by American Chemical Society (ACS) in Analytical Chemistry
- Vol. 77 (24) , 7878-7886
- https://doi.org/10.1021/ac050781s
Abstract
This paper reports a protocol that improves the resolving power of isoelectric focusing (IEF) in a polymeric microfluidic chip. This method couples several stages of IEF in series by first focusing proteins in a straight channel using broad-range ampholytes and then refocusing segments of the first channel into secondary channels that branch from the first one at T-junctions. Experiments demonstrate that several fluorescent proteins that had focused within a segment of the straight channel in the first stage were refocused at significantly higher resolution due to the shallower pH gradient and higher electrical field gradient. Two variants of green fluorescent protein from the second-stage IEF fractionation were further separated in a third stage. Three stages of IEF were completed in less than 25 min at electric field strengths ranging from 50 to 214 V/cm.Keywords
This publication has 21 references indexed in Scilit:
- Poly‐N‐hydroxyethylacrylamide as a novel, adsorbed coating for protein separation by capillary electrophoresisElectrophoresis, 2003
- Monolithic membrane valves and diaphragm pumps for practical large-scale integration into glass microfluidic devicesSensors and Actuators B: Chemical, 2003
- Monolithic Microfabricated Valves and Pumps by Multilayer Soft LithographyScience, 2000
- Computer Simulations of Electrokinetic Transport in Microfabricated Channel StructuresAnalytical Chemistry, 1998
- Fractionation of carrier ampholytes in multicompartment electrolyzers with isoelectric membranesElectrophoresis, 1995
- Micromachining a Miniaturized Capillary Electrophoresis-Based Chemical Analysis System on a ChipScience, 1993
- Computer simulation and experimental validation of the electrophoretic behavior of proteins. 2. Model improvement and application to isotachophoresisAnalytical Chemistry, 1992
- Computer simulation and experimental validation of the electrophoretic behavior of proteinsAnalytical Chemistry, 1989
- Effect of temperature gradients on the efficiency of capillary zone electrophoresis separationsAnalytical Chemistry, 1989
- Isoelectric Fractionation, Analysis, and Characterization of Ampholytes in Natural pH Gradients. IV. Further Studies on the Resolving Power in Connection with Separation of Myoglobins.Acta Chemica Scandinavica, 1966