Simplified 2-D CE-MS mapping: Analysis of proteolytic digests
It has been demonstrated that CE‐MS is a very useful hyphenated technique for proteomic studies. However, the huge amount of data stored in a single CE‐MS run makes it necessary to account with procedures able to extract all the relevant information made available by CE‐MS. In this work, we present...
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Published in | Electrophoresis Vol. 28; no. 9; pp. 1335 - 1344 |
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Main Authors | , |
Format | Journal Article |
Language | English |
Published |
Weinheim
WILEY-VCH Verlag
01.05.2007
WILEY‐VCH Verlag |
Subjects | |
Online Access | Get full text |
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Summary: | It has been demonstrated that CE‐MS is a very useful hyphenated technique for proteomic studies. However, the huge amount of data stored in a single CE‐MS run makes it necessary to account with procedures able to extract all the relevant information made available by CE‐MS. In this work, we present a new and easy approach capable of generating a simplified 2‐D map from CE‐MS raw data. This new approach provides the automatic detection and characterization of the most abundant ions from the CE‐MS data including their mass‐to‐charge (m/z) values, ion intensities and analysis times. It is demonstrated that visualization of CE‐MS data in this simplified 2‐D format allows: (i) an easy and simultaneous visual inspection of large datasets, (ii) an immediate perception of relevant differences in closely related samples, (iii) a rapid monitoring of data quality levels in different samples, and (iv) a fast discrimination between comigrating polypeptides and ESI‐MS fragmentation ions. The strategy proposed in this work does not rely on an excellent mass accuracy for peak detection and filtering, since MS values obtained from an IT analyzer are used. Moreover, the methodology developed works directly with the CE‐MS raw data, without interference by the user, giving simultaneously a simplified 2‐D map and a much easier and more complete data evaluation. Besides, this procedure can easily be implemented in any CE‐MS laboratory. The usefulness of this approach is validated by studying the very similar trypsin digests from bovine, rabbit and horse cytochrome c. It is demonstrated that this simplified 2‐D approach allows specific markers for each species to be obtained in a fast and simple way. |
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Bibliography: | istex:2838BCFB34005CEB163F69004F97B91C35D938B8 Comunidad Autonoma de Madrid - No. S-505/AGR-0153 Ministerio de Educacion y Ciencia - No. AGL2005-05320-C02-01 ark:/67375/WNG-Z6MPL95S-W ArticleID:ELPS200600357 ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 ObjectType-Article-1 ObjectType-Feature-2 |
ISSN: | 0173-0835 1522-2683 |
DOI: | 10.1002/elps.200600357 |