Microfluidic device having an immobilized pH gradient and page gels for protein separation and analysis
| DWPI Title: Two-dimensional separation of proteins and related polypeptide molecules involves forming isoelectric focusing stage within microchannel of microfluidic device by introducing acrylamide/bisacrylamide solution into length of microchannel |
| 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: The method is useful for two-dimensional separation of proteins and related polypeptide molecules (claimed). |
| Advantage: The method allows rapid and simple two-dimensional separation of proteins and related polypeptide molecules with high resolution and without using handling gel strips. |
| Novelty: The method comprises forming an isoelectric focusing (IEF) stage within a first microchannel of a microfluidic device. The microfluidic device comprises branching side microchannels (108) present in fluid communication with the first microchannel. The first microchannel and each of the branching side microchannels comprise interior surfaces comprising an acrylate-terminated coating, and have a length and a width. The step of forming the IEF stage comprises introducing an acrylamide/bisacrylamide solution into a length of the first microchannel and forming a pH gradient. |
| Filed: 11/2/2012 |
| Application Number: US13667254A |
| Tech ID: SD 11177.1 |
| 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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