| LENČO, Juraj, Siddharth JADEJA, Denis K. NAPLEKOV, Oleg V. KROKHIN, Maria A. KHALIKOVA, Petr NOVÁKOVÁ, Jiří URBAN, Ken BROECKHOVEN, Lucie NOVÁKOVÁ a František ŠVEC. Reversed-Phase Liquid Chromatography of Peptides for Bottom-Up Proteomics: A Tutorial. Journal of Proteome Research. Washington, USA: American Chemical Society, 2022, roč. 21, č. 12, s. 2846-2892. ISSN 1535-3893. Dostupné z: https://dx.doi.org/10.1021/acs.jproteome.2c00407. |
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@article{2249090,
author = {Lenčo, Juraj and Jadeja, Siddharth and Naplekov, Denis K. and Krokhin, Oleg V. and Khalikova, Maria A. and Nováková, Petr and Urban, Jiří and Broeckhoven, Ken and Nováková, Lucie and Švec, František},
article_location = {Washington, USA},
article_number = {12},
doi = {http://dx.doi.org/10.1021/acs.jproteome.2c00407},
keywords = {bottom-up proteomics; peptide separation; reversed-phase liquid chromatography; peptide properties; method development; protein separation; tutorial},
language = {eng},
issn = {1535-3893},
journal = {Journal of Proteome Research},
title = {Reversed-Phase Liquid Chromatography of Peptides for Bottom-Up Proteomics: A Tutorial},
url = {https://pubs.acs.org/doi/10.1021/acs.jproteome.2c00407},
volume = {21},
year = {2022}
}
TY - JOUR
ID - 2249090
AU - Lenčo, Juraj - Jadeja, Siddharth - Naplekov, Denis K. - Krokhin, Oleg V. - Khalikova, Maria A. - Nováková, Petr - Urban, Jiří - Broeckhoven, Ken - Nováková, Lucie - Švec, František
PY - 2022
TI - Reversed-Phase Liquid Chromatography of Peptides for Bottom-Up Proteomics: A Tutorial
JF - Journal of Proteome Research
VL - 21
IS - 12
SP - 2846-2892
EP - 2846-2892
PB - American Chemical Society
SN - 15353893
KW - bottom-up proteomics
KW - peptide separation
KW - reversed-phase liquid chromatography
KW - peptide properties
KW - method development
KW - protein separation
KW - tutorial
UR - https://pubs.acs.org/doi/10.1021/acs.jproteome.2c00407
N2 - The performance of the current bottom-up liquid chromatography hyphenated with mass spectrometry (LC-MS) analyses has undoubtedly been fueled by spectacular progress in mass spectrometry. It is thus not surprising that the MS instrument attracts the most attention during LC-MS method development, whereas optimizing conditions for peptide separation using reversed-phase liquid chromatography (RPLC) remains somewhat in its shadow. Consequently, the wisdom of the fundaments of chromatography is slowly vanishing from some laboratories. However, the full potential of advanced MS instruments cannot be achieved without highly efficient RPLC. This is impossible to attain without understanding fundamental processes in the chromatographic system and the properties of peptides important for their chromatographic behavior. We wrote this tutorial intending to give practitioners an overview of critical aspects of peptide separation using RPLC to facilitate setting the LC parameters so that they can leverage the full capabilities of their MS instruments. After briefly introducing the gradient separation of peptides, we discuss their properties that affect the quality of LC-MS chromatograms the most. Next, we address the in-column and extra-column broadening. The last section is devoted to key parameters of LC-MS methods. We also extracted trends in practice from recent bottom-up proteomics studies and correlated them with the current knowledge on peptide RPLC separation.
ER -
LENČO, Juraj, Siddharth JADEJA, Denis K. NAPLEKOV, Oleg V. KROKHIN, Maria A. KHALIKOVA, Petr NOVÁKOVÁ, Jiří URBAN, Ken BROECKHOVEN, Lucie NOVÁKOVÁ a František ŠVEC. Reversed-Phase Liquid Chromatography of Peptides for Bottom-Up Proteomics: A Tutorial. \textit{Journal of Proteome Research}. Washington, USA: American Chemical Society, 2022, roč.~21, č.~12, s.~2846-2892. ISSN~1535-3893. Dostupné z: https://dx.doi.org/10.1021/acs.jproteome.2c00407.
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