Highly efficient peptide separations in proteomics Part 1. Unidimensional high performance liquid chromatography

Sample complexity and dynamic range constitute enormous challenges in proteome analysis. The back-end technology in typical proteomics platforms, namely mass spectrometry (MS), can only tolerate a certain complexity, has a limited dynamic range per spectrum and is very sensitive towards ion suppress...

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Bibliographic Details
Published in:Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2008-04, Vol.866 (1-2), p.48-63
Main Authors: SANDRA, Koen, MOSHIR, Mahan, D'HONDT, Filip, VERLEYSEN, Katleen, KAS, Koen, SANDRA, Pat
Format: Article
Language:English
Subjects:
Analysis
Analytical, structural and metabolic biochemistry
Biological and medical sciences
Chromatography, High Pressure Liquid - methods
Fundamental and applied biological sciences. Psychology
General pharmacology
Medical sciences
Peptides - isolation & purification
Pharmacology. Drug treatments
Proteomics
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Online Access:Get full text
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Summary:Sample complexity and dynamic range constitute enormous challenges in proteome analysis. The back-end technology in typical proteomics platforms, namely mass spectrometry (MS), can only tolerate a certain complexity, has a limited dynamic range per spectrum and is very sensitive towards ion suppression. Therefore, component overlap has to be minimized for successful mass spectrometric analysis and subsequent protein identification and quantification. The present review describes the advances that have been made in liquid-based separation techniques with focus on the recent developments to boost the resolving power. The review is divided in two parts; the first part deals with unidimensional liquid chromatography and the second part with bi- and multidimensional liquid-based separation techniques. Part 1 mainly focuses on reversed-phase HPLC due to the fact that it is and will, in the near future, remain the technique of choice to be hyphenated with MS. The impact of increasing the column length, decreasing the particle diameter, replacing the traditional packed beds by monolithics, amongst others, is described. The review is complemented with data obtained in the laboratories of the authors.
ISSN:1570-0232
1873-376X
DOI:10.1016/j.jchromb.2007.10.034