High-performance nano-flow liquid chromatography column combined with high- and low-collision energy data-independent acquisition enables targeted and discovery identification of modified ribonucleotides by mass spectrometry

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• dc.contributor.author Espadas, Guadalupe
• dc.contributor.author Morales Sanfrutos, Julia
• dc.contributor.author Medina, Rebeca
• dc.contributor.author Lucas, Morghan C.
• dc.contributor.author Novoa, Eva Maria
• dc.contributor.author Sabidó Aguadé, Eduard, 1981-
• dc.date.accessioned 2022-04-04T06:31:17Z
• dc.date.available 2022-04-04T06:31:17Z
• dc.date.issued 2022
• dc.description.abstract Over 170 post-transcriptional RNA modifications have been described and are common in all kingdoms of life. These modifications range from methylation to complex chemical structures, with methylation being the most abundant. RNA modifications play a key role in RNA folding and function and their dysregulation in humans has been linked to several diseases such as cancer, metabolic diseases or neurological disorder. Nowadays, liquid chromatography-tandem mass spectrometry is considered the gold standard method for the identification and quantification of these modifications due to its sensitivity and accuracy. However, the analysis of modified ribonucleosides by mass spectrometry is complex due to the presence of positional isomers. In this scenario, optimal separation of these compounds by highly sensitive liquid chromatography combined with the generation of high-information spectra is critical to unequivocally identify them, especially in high-complex mixtures. Here we present an analytical method that comprises a new type of mixed-mode nano-flow liquid chromatography column combined with high- and low-collision energy data-independent mass spectrometric acquisition for the identification and quantitation of modified ribonucleosides. The method produces content-rich spectra and combines targeted and screening capabilities thus enabling the identification of a variety of modified nucleosides in biological matrices by single-shot liquid chromatographic analysis coupled to mass spectrometry.
• dc.description.sponsorship The CRG/UPF Proteomics Unit is part of the Spanish Infrastructure for Omics Technologies (ICTS OmicsTech) and it is a member of the ProteoRed PRB3 consortium, which is supported by grant PT17/0019 of the PE I+D+i 2013–2016 from the Instituto de Salud Carlos III (ISCIII) and ERDF. We acknowledge support from the Spanish Ministry of Science and Innovation (projects CTQ2016–80364-P to ES and PGC2018–098152-A-100 to EMN) and “Centro de Excelencia Severo Ochoa 2013–2017″, SEV-2012–0208; and from “Secretaria d'Universitats i Recerca del Departament d'Economia i Coneixement de la Generalitat de Catalunya” (2017SGR595). We also acknowledge support of the Spanish Ministry of Science and Innovation to the EMBL partnership, the Centro de Excelencia Severo Ochoa and the CERCA Programme / Generalitat de Catalunya.”
• dc.format.mimetype application/pdf
• dc.identifier.citation Espadas G, Morales-Sanfrutos J, Medina R, Lucas MC, Novoa EM, Sabidó E. High-performance nano-flow liquid chromatography column combined with high- and low-collision energy data-independent acquisition enables targeted and discovery identification of modified ribonucleotides by mass spectrometry. J Chromatogr A. 2022 Feb 22;1665:462803. DOI: 10.1016/j.chroma.2022.462803
• dc.identifier.doi http://dx.doi.org/10.1016/j.chroma.2022.462803
• dc.identifier.issn 0021-9673
• dc.identifier.uri http://hdl.handle.net/10230/52821
• dc.language.iso eng
• dc.publisher Elsevier
• dc.relation.ispartof J Chromatogr A. 2022 Feb 22;1665:462803
• dc.relation.projectID info:eu-repo/grantAgreement/ES/1PE/CTQ2016–80364-P
• dc.relation.projectID info:eu-repo/grantAgreement/ES/2PE/PGC2018–098152-A-100
• dc.rights © 2022 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/)
• dc.rights.accessRights info:eu-repo/semantics/openAccess
• dc.rights.uri http://creativecommons.org/licenses/by/4.0/
• dc.subject.keyword Chromatography
• dc.subject.keyword Data-independent acquisition
• dc.subject.keyword Epitranscriptomics
• dc.subject.keyword Mass spectrometry
• dc.subject.keyword RNA
• dc.subject.keyword Ribonucleosides
• dc.title High-performance nano-flow liquid chromatography column combined with high- and low-collision energy data-independent acquisition enables targeted and discovery identification of modified ribonucleotides by mass spectrometry
• dc.type info:eu-repo/semantics/article
• dc.type.version info:eu-repo/semantics/publishedVersion