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Micro-proteome analysis using micro-chromatofocusing in intact protein separations.

ABSTRACT: Multi-dimensional liquid-based separation is required for fractionation and mapping of complex protein mixtures from cells. A method that has been used as the first dimension in such separations is chromatofocusing (CF), which is based on generating a pH gradient on an anion exchange column. The use of pH in the first dimension is essential where pH is a fundamental property of proteins and can provide information on post-translationally modified forms of a protein. In this work, a micro-chromatofocusing technique is introduced which can separate microgram levels of proteins from cell lysates for further analysis by LC-MS/MS. It is shown that this method can analyze 10 microg of sample and detect nearly 700-800 proteins from ovarian cancer cell line lysates.

SUBMITTER: Kim H

PROVIDER: S-EPMC2479787 | biostudies-literature | 2008 Jun

REPOSITORIES: biostudies-literature

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Micro-proteome analysis using micro-chromatofocusing in intact protein separations.

Kim Hyeyeung H Lubman David M DM

Journal of chromatography. A 20080327 1

Multi-dimensional liquid-based separation is required for fractionation and mapping of complex protein mixtures from cells. A method that has been used as the first dimension in such separations is chromatofocusing (CF), which is based on generating a pH gradient on an anion exchange column. The use of pH in the first dimension is essential where pH is a fundamental property of proteins and can provide information on post-translationally modified forms of a protein. In this work, a micro-chromat ...[more]

PMID: 18407281

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