High-throughput and high-resolution two dimensional mapping of pI and m/z using a microchip in a matrix-assisted laser desorption/ionization time-of-flight mass spectrometer |
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Authors: | Fujita Machiko Hattori Wataru Sano Tohru Baba Masakazu Someya Hiroko Miyazaki Kenji Kamijo Ken'ichi Takahashi Katsutoshi Kawaura Hisao |
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Institution: | Fundamental and Environmental Research laboratories, NEC Corporation, Tsukuba, Ibaraki, Japan. m-fujita@frl.cl.nec.co.jp |
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Abstract: | We have developed a high-throughput, two-dimensional-mapping (isoelectric point pI], mass-to-charge ratio m/z]) method by combining a capillary isoelectric focusing chip sealed with removable resin tape and a matrix-assisted laser desorption/ionization time-of-flight mass spectrometer. Sample proteins are separated in a meandering channel on the chip and immediately frozen. The tape is then removed and the proteins are freeze-dried. The freeze-drying maintains the separation state of the proteins and prevents movement of the sample solution, which can reduce pI resolution. A matrix solution is then applied and mass spectrometry is carried out by laser irradiation. The whole process takes less than 70 min, more than 10 times faster than with two-dimensional, polyacrylamide gel electrophoresis. |
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