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  Nanoflow liquid chromatography coupled to matrix-assisted laser desorption/ionization mass spectrometry: Sample preparation, data analysis, and application to the analysis of complex peptide mixtures

Mirgorodskaya, E., Braeuer, C., Fucini, P., Lehrach, H., & Gobom, J. (2005). Nanoflow liquid chromatography coupled to matrix-assisted laser desorption/ionization mass spectrometry: Sample preparation, data analysis, and application to the analysis of complex peptide mixtures. PROTEOMICS, 5(2), 399-408. doi:10.1002/pmic.200400984.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0010-871D-8 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0010-871E-6
Genre: Journal Article

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Mirgorodskaye et al. - Proteomics.pdf (Any fulltext), 407KB
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Mirgorodskaye et al. - Proteomics.pdf
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 Creators:
Mirgorodskaya, Ekaterina1, Author
Braeuer, Corina1, Author
Fucini, Paola2, Author              
Lehrach, Hans3, Author              
Gobom, Johan3, Author              
Affiliations:
1Max Planck Society, ou_persistent13              
2Ribosomes, Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1433558              
3Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1433550              

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Free keywords: Liquid chromatography, Matrix-assisted laser desorption/ionisation; Peptides; Protein identification; Tandem mass spectrometry
 Abstract: We report the development of a robust interface for off-line coupling of nano liquid chromatography (LC) to matrix-assisted laser desorption/ionisation-mass spectrometry (MALDI-MS) and its application to the analysis of proteolytic digests of proteins, both isolated and in mixtures. The interface makes use of prestructured MALDI sample supports to concentrate the effluent to a small sample plate area and localize the MALDI sample to a predefined array, thereby enriching the analyte molecules and facilitating automated MALDI-MS analysis. Parameters that influence the preparation of MALDI samples from the LC effluent were evaluated with regard to detection sensitivity, spectra quality, and reproducibility of the method. A procedure for data processing is described. The presented nano LC MALDI-MS system allowed the detection of several peptides from a tryptic digest of bovine serum albumin, at analyzed amounts corresponding to one femtomole of the digested protein. For the identification of native proteins isolated from mouse brain by two-dimensional gel electrophoresis, nano LC MALDI-MS increased the number of detected peptides, thereby allowing identification of proteins that could not be identified by direct MALDI-MS analysis. The ability to identify proteins in complex mixtures was evaluated for the analysis of Escherichia coli 50S ribosomal subunit. Out of the 33 expected proteins, 30 were identified by MALDI tandem time of flight fragment ion fingerprinting.

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Language(s): eng - English
 Dates: 2005-01-13
 Publication Status: Published in print
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 Identifiers: eDoc: 272954
DOI: 10.1002/pmic.200400984
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Title: PROTEOMICS
Source Genre: Journal
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Pages: - Volume / Issue: 5 (2) Sequence Number: - Start / End Page: 399 - 408 Identifier: ISSN: 1615-9853