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Enhanced sequence coverage in tryptic fragment analysis by two-dimensional HPLC/MS using a monolithic silica capillary column.

Hironobu Morisaka·Koji Hata·Joji Mima·Tetsuya Tanigawa·Masahiro Furuno·Norio Ishizuka·Nobuo Tanaka·Mitsuyoshi Ueda
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Abstract

The HPLC/MS system, in which a monolithic silica capillary column is directly connected to an electronspray-ionization mass spectrometer, showed superior performance at high mobile phase linear velocity. A two-dimensional (2D) HPLC/MS system was established, using an ion-exchange particle-packed capillary column at the first dimension and a monolithic silica capillary column at the second dimension. In an analysis of tryptic fragments from bovine serum albumin, an 81% sequence coverage, obtained by the 2D-HPLC/MS system, increased by 23% as compared to a 1D-HPLC/MS system. This 2D-HPLC/MS system using a monolithic silica capillary column should be useful for enhancing sequence coverage of tryptic fragments in proteomics.

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Amino Acid SequenceChromatography, High Pressure LiquidChromatography, Ion ExchangeMolecular Sequence DataProteomicsSequence Analysis, ProteinSerum Albumin, BovineSpectrometry, Mass, Electrospray IonizationTrypsinJournal ArticleResearch Support, Non-U.S. Gov't

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Hironobu Morisaka, Koji Hata, Joji Mima, Tetsuya Tanigawa, Masahiro Furuno, Norio Ishizuka, Nobuo Tanaka, Mitsuyoshi Ueda. 2006. Enhanced sequence coverage in tryptic fragment analysis by two-dimensional HPLC/MS using a monolithic silica capillary column.. https://doi.org/10.1271/bbb.60115

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