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Bioinertization of NanoLC/MS/MS Systems by Depleting Metal Ions From the Mobile Phases for Phosphoproteomics
We have successfully developed a bioinertized nanoflow LC/MS/MS (nanoLC/MS/MS) system for the highly sensitive analysis of phosphopeptides by depleting metal ions from the mobile phase. We found that not only direct contact of phosphopeptides with metal components, but also indirect contact with nan...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Biochemistry and Molecular Biology
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10172917/ https://www.ncbi.nlm.nih.gov/pubmed/36958626 http://dx.doi.org/10.1016/j.mcpro.2023.100535 |
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author | Komori, Yumi Niinae, Tomoya Imami, Koshi Yanagibayashi, Jun Yasunaga, Kenichi Imamura, Shinya Tomita, Masami Ishihama, Yasushi |
author_facet | Komori, Yumi Niinae, Tomoya Imami, Koshi Yanagibayashi, Jun Yasunaga, Kenichi Imamura, Shinya Tomita, Masami Ishihama, Yasushi |
author_sort | Komori, Yumi |
collection | PubMed |
description | We have successfully developed a bioinertized nanoflow LC/MS/MS (nanoLC/MS/MS) system for the highly sensitive analysis of phosphopeptides by depleting metal ions from the mobile phase. We found that not only direct contact of phosphopeptides with metal components, but also indirect contact with nanoLC pumps through the mobile phase causes significant losses during the recovery of phosphopeptides. Moreover, electrospray ionization was adversely affected by the mobile phase containing multiple metal ions as well as by the sample solvents contaminated with metal ions used in immobilized metal ion affinity chromatography for phosphopeptide enrichment. To solve these problems, metal ions were depleted by inserting an online metal ion removal device containing metal-chelating membranes between the gradient mixer and the autosampler. As a result, the peak areas of the identified phosphopeptides increased an average of 9.9-fold overall and 77-fold for multiply phosphorylated peptides with the insertion of the online metal ion removal system. This strategy would be applicable to the highly sensitive analysis of other phosphorylated biomolecules by microscale-LC/MS/MS. |
format | Online Article Text |
id | pubmed-10172917 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-101729172023-05-12 Bioinertization of NanoLC/MS/MS Systems by Depleting Metal Ions From the Mobile Phases for Phosphoproteomics Komori, Yumi Niinae, Tomoya Imami, Koshi Yanagibayashi, Jun Yasunaga, Kenichi Imamura, Shinya Tomita, Masami Ishihama, Yasushi Mol Cell Proteomics Technological Innovation and Resources We have successfully developed a bioinertized nanoflow LC/MS/MS (nanoLC/MS/MS) system for the highly sensitive analysis of phosphopeptides by depleting metal ions from the mobile phase. We found that not only direct contact of phosphopeptides with metal components, but also indirect contact with nanoLC pumps through the mobile phase causes significant losses during the recovery of phosphopeptides. Moreover, electrospray ionization was adversely affected by the mobile phase containing multiple metal ions as well as by the sample solvents contaminated with metal ions used in immobilized metal ion affinity chromatography for phosphopeptide enrichment. To solve these problems, metal ions were depleted by inserting an online metal ion removal device containing metal-chelating membranes between the gradient mixer and the autosampler. As a result, the peak areas of the identified phosphopeptides increased an average of 9.9-fold overall and 77-fold for multiply phosphorylated peptides with the insertion of the online metal ion removal system. This strategy would be applicable to the highly sensitive analysis of other phosphorylated biomolecules by microscale-LC/MS/MS. American Society for Biochemistry and Molecular Biology 2023-03-22 /pmc/articles/PMC10172917/ /pubmed/36958626 http://dx.doi.org/10.1016/j.mcpro.2023.100535 Text en © 2023 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Technological Innovation and Resources Komori, Yumi Niinae, Tomoya Imami, Koshi Yanagibayashi, Jun Yasunaga, Kenichi Imamura, Shinya Tomita, Masami Ishihama, Yasushi Bioinertization of NanoLC/MS/MS Systems by Depleting Metal Ions From the Mobile Phases for Phosphoproteomics |
title | Bioinertization of NanoLC/MS/MS Systems by Depleting Metal Ions From the Mobile Phases for Phosphoproteomics |
title_full | Bioinertization of NanoLC/MS/MS Systems by Depleting Metal Ions From the Mobile Phases for Phosphoproteomics |
title_fullStr | Bioinertization of NanoLC/MS/MS Systems by Depleting Metal Ions From the Mobile Phases for Phosphoproteomics |
title_full_unstemmed | Bioinertization of NanoLC/MS/MS Systems by Depleting Metal Ions From the Mobile Phases for Phosphoproteomics |
title_short | Bioinertization of NanoLC/MS/MS Systems by Depleting Metal Ions From the Mobile Phases for Phosphoproteomics |
title_sort | bioinertization of nanolc/ms/ms systems by depleting metal ions from the mobile phases for phosphoproteomics |
topic | Technological Innovation and Resources |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10172917/ https://www.ncbi.nlm.nih.gov/pubmed/36958626 http://dx.doi.org/10.1016/j.mcpro.2023.100535 |
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