Journal
JOURNAL OF CHROMATOGRAPHY A
Volume 1111, Issue 2, Pages 200-205Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.chroma.2005.11.117
Keywords
chip technology; two-dimensional separations; proteins; capillary isoelectric focusing; matrix-assisted laser desorption/ionization time-of-flight mass; spectrometer (MALDI-TOF-MS)
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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 taster than with two-dimensional, polyacrylamide gel electrophoresis. (c) 2005 Elsevier B.V. All rights reserved.
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