Microfluidic device having an immobilized pH gradient and PAGE gels for protein separation and analysis
| DWPI Title: Microchannel device comprises isoelectric focusing stage having central microchannel including surface(s) with acrylate-terminated coating and pH gradient polymer gel, and orthogonal polyacrylamide electrophoretic gel separation stage |
| Abstract: Disclosed is a novel microfluidic device enabling on-chip implementation of a two-dimensional separation methodology. Previously disclosed microscale immobilized pH gradients (IPG) are combined with perpendicular polyacrylamide gel electrophoresis (PAGE) microchannels to achieve orthogonal separations of biological samples. Device modifications enable inclusion of sodium dodecyl sulfate (SDS) in the second dimension. The device can be fabricated to use either continuous IPG gels, or the microscale isoelectric fractionation membranes we have also previously disclosed, for the first dimension. The invention represents the first all-gel two-dimensional separation microdevice, with significantly higher resolution power over existing devices. |
| Use: As two-dimensional microchannel separation device (claimed). |
| Advantage: The two-dimensional microchannel separation device provides simple method for providing 2-D separation of molecular species in continuous, all-gel format without need for handling gel strips, by providing separation dimensions (IEF and PAGE) in continuous gel format, where first dimension has immobilized pH gradient for rapid and high resolution (IEF), and second dimension has several microchannels for polyacrylamide gel electrophoresis (PAGE). The device provides integrated, miniaturized 2-D IEF/PAGE analysis device provided in microfluidic format. The device further allows for introducing anionic surfactant such as sodium dodecyl sulfate (SDS) into second dimension. |
| Novelty: Two-dimensional microchannel separation device (100) comprises: isoelectric focusing (IEF) stage in communication with orthogonal polyacrylamide electrophoretic gel (PAGE) separation stage, where IEF stage comprises central microchannel (102) including interior surface(s) with acrylate-terminated coating, and immobilized pH gradient polymer gel bonded to coating, partially filling microchannel; electrode at each of two ends of IEF stage and PAGE stage; power supply; and switching device for applying electric potential between any two of electrodes. |
| Filed: 8/31/2009 |
| Application Number: US2009551047A |
| Tech ID: SD 11177.0 |
| This invention was made with Government support under Contract No. DE-NA0003525 awarded by the United States Department of Energy/National Nuclear Security Administration. The Government has certain rights in the invention. |
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