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MALDI/MS Comparison of Fe-NTA Immobilized Metal Affinity Chromatography and Commercially-Available Metal Oxide Affinity Resins for Phosphopeptide Enrichment

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Applications of Mass Spectrometry in Life Safety

Abstract

Immobilized metal ion affinity chromatography in combination with mass spectrometry has been used to determine the extent of phosphorylation and the specific sites of phosphorylation on proteins. There are several advantages to this combined approach. Immobilized metal ion affinity chromatography is the use of resins with metal constituents and metal oxides to enrich and isolate phosphopeptides. By selectively enriching phosphopeptides on media prior to MS analyses, suppression effects can be greatly reduced. In this report, we have investigated several resins from various sources to assess the phosphopeptide enrichment capabilities of each prior to mass spectrometric analyses. The phosphopeptide enrichment capabilities of six different resins, Glygen TiO2, ZrO2, and mixed ZrO2 and TiO2 NuTips, Titansphere TiO2 resin, PhosTrap magnetic titanium beads, and a Fe-NTA resin, are compared.

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Authors and Affiliations

  1. Laboratory of Structural Biology, National Institute of Environmental Health Sciences, National Institutes of Health, PO Box 12233, MD F0-03, Research Triangle Park, NC, 27709, USA

    Matthew B. Gates, Kenneth B. Tomer & Leesa J. Deterding

Authors
  1. Matthew B. Gates
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  2. Kenneth B. Tomer
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  3. Leesa J. Deterding
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Editor information

Editors and Affiliations

  1. DWI e.V. and Institute of Technical and Macromolecular Chemistry, RWTH Aachen University, Germany

    Crisan Popescu

  2. University “Aurel Vlaicu”, Arad, Romania

    Alina D. Zamfir  & Nicolae Dinca  & 

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Gates, M.B., Tomer, K.B., Deterding, L.J. (2008). MALDI/MS Comparison of Fe-NTA Immobilized Metal Affinity Chromatography and Commercially-Available Metal Oxide Affinity Resins for Phosphopeptide Enrichment. In: Popescu, C., Zamfir, A.D., Dinca, N. (eds) Applications of Mass Spectrometry in Life Safety. NATO Science for Peace and Security Series A: Chemistry and Biology. Springer, Dordrecht. https://doi.org/10.1007/978-1-4020-8811-7_3

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