Qiu, W., Evans, C.A. orcid.org/0000-0003-4356-9216, Landels, A. et al. (2 more authors) (2020) Phosphopeptide enrichment for phosphoproteomic analysis - a tutorial and review of novel materials. Analytica Chimica Acta, 1129. pp. 158-180. ISSN 0003-2670
Abstract
Significant technical advancements in phosphopeptide enrichment have enabled the identification of thousands of p-peptides (mono and multiply phosphorylated) in a single experiment. However, it is still not possible to enrich all p-peptide species in a single step. A range of new techniques and materials has been developed, with the potential to provide a step-change in phosphopeptide enrichment. The first half of this review contains a tutorial for new potential phosphoproteomic researchers; discussing the key steps of a typical phosphoproteomic experiment used to investigate canonical phosphorylation sites (serine, threonine and tyrosine). The latter half then show-cases the latest developments in p-peptide enrichment including: i) Strategies to mitigate non-specific binding in immobilized metal ion affinity chromatography and metal oxide affinity chromatography protocols; ii) Techniques to separate multiply phosphorylated peptides from monophosphorylated peptides (including canonical from non-canonical phosphorylated peptides), or to simultaneously co-enrich other post-translational modifications; iii) New hybrid materials and methods directed towards enhanced selectivity and efficiency of metal-based enrichment; iv) Novel materials that hold promise for enhanced phosphotyrosine enrichment. A combination of well-understood techniques and materials is much more effective than any technique in isolation; but the field of phosphoproteomics currently requires benchmarking of novel materials against current methodologies to fully evaluate their utility in peptide based proteoform analysis.
Metadata
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Copyright, Publisher and Additional Information: | © 2020 Elsevier. This is an author produced version of a paper subsequently published in Analytica Chimica Acta. Uploaded in accordance with the publisher's self-archiving policy. Article available under the terms of the CC-BY-NC-ND licence (https://creativecommons.org/licenses/by-nc-nd/4.0/). | ||||||
Keywords: | Phosphoproteoform; Canonical and non-canonical phosphorylation; Enrichment; Optimization; Phosphoproteomics; Phosphopeptide | ||||||
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Institution: | The University of Sheffield | ||||||
Academic Units: | The University of Sheffield > Faculty of Engineering (Sheffield) > Department of Chemical and Biological Engineering (Sheffield) | ||||||
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Depositing User: | Symplectic Sheffield | ||||||
Date Deposited: | 15 Jul 2020 06:46 | ||||||
Last Modified: | 24 May 2022 12:07 | ||||||
Status: | Published | ||||||
Publisher: | Elsevier | ||||||
Refereed: | Yes | ||||||
Identification Number: | https://doi.org/10.1016/j.aca.2020.04.053 |