Proteomics

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Bioinertization of nanoLC/MS/MS systems by depleting metal ions from the mobile phases for phosphoproteomics

ABSTRACT: We have successfully bioinertized a nanoLC/MS/MS system for the highly sensitive analysis of phosphopeptides by depleting metal ions from the mobile phase. We found that not only the direct contact of phosphopeptides with metal materials, but also the indirect contact with HPLC pump through the mobile phase causes the significant loss in the recovery of phosphopeptides. Moreover, we observed that electrospray ionization was affected by the mobile phase containing multiple metal ions as well as the sample solutions contaminated with metal ions used by immobilized metal ion affinity chromatography for the phosphopeptide enrichment. To solve these problems, metal ions were depleted by inserting an on-line filter device containing metal chelating resins between the gradient mixer and the autosampler. As a result, we were able to increase the identification number of phosphopeptides by a factor of 1.5. This strategy would be applicable to highly sensitive analysis of other phosphorylated biomolecules by nanoLC/MS/MS.

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Yasushi Ishihama 

PROVIDER: PXD034942 | JPOST Repository | Fri Mar 24 00:00:00 GMT 2023

REPOSITORIES: jPOST

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180120TN_02_Ctrl.mgf Mgf
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180120TN_04_Ti2_1.mgf Mgf
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Bioinertization of NanoLC/MS/MS Systems by Depleting Metal Ions From the Mobile Phases for Phosphoproteomics.

Komori Yumi Y   Niinae Tomoya T   Imami Koshi K   Yanagibayashi Jun J   Yasunaga Kenichi K   Imamura Shinya S   Tomita Masami M   Ishihama Yasushi Y  

Molecular & cellular proteomics : MCP 20230322 5

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 me  ...[more]

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