Specific Enrichment of Phosphoproteins Using Functionalized Multivalent Nanoparticles
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Analysis of protein phosphorylation remains a significant challenge due to the low abundance of phosphoproteins and the low stoichiometry of phosphorylation, which requires effective enrichment of phosphoproteins. Here we have developed superparamagnetic nanoparticles (NPs) whose surface is functionalized by multivalent ligand molecules that specifically bind to the phosphate groups on any phosphoproteins. These NPs enrich phosphoproteins from complex cell and tissue lysates with high specificity as confirmed by SDS-PAGE analysis with a phosphoprotein-specific stain and mass spectrometry analysis of the enriched phosphoproteins. This method enables universal and effective capture, enrichment, and detection of intact phosphoproteins toward a comprehensive analysis of the phosphoproteome.
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Intact proteoform enrichment necessary for top-down proteomics is relatively underdeveloped.
“On the other hand, intact proteoform enrichment, which is necessary for top-down proteomics, is relatively underdeveloped [56].”
From Top-down proteomics: challenges, innovations, and applications in basic and clinical research (2020) · cites this paper as [56]
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Ortiz‐Gómez, I., & Calderón‐Celis, F. (2026) · Advances in experimental medicine and biology · doi:10.1007/978-3-032-21638-0_3
Advances in Sample Preparation for Protein Top-Down Mass Spectrometry
Zhang, M., Xue, J.-Y., Yang, S.-R., Yu, Y.-L., Liu, Z.-Y., & Wang, F.-J. (2026) · Analysis and Testing Technology and Instruments · doi:10.3724/j.1006-3757.2026.05.004
Quantitative Top-Down Proteomics: A Review.
Cupp-Sutton, K. A., Atallah, M. A., Zhao, Z.-T., Anjum, S., Chowdhury, T., Sunny, A., Szamweber, P., & Wu, S. (2026) · Mass spectrometry reviews (Print) · doi:10.1002/mas.70051
- Authors
- Hwang, L., Ayaz-Guner, S., Gregorich, Z. R., Cai, W., Valeja, S. G., Jin, S., & Ge, Y.
- Affiliations
- University of Wisconsin–Madison (US)
- Department of Chemistry, ‡Department of Cell and Regenerative Biology, §Molecular and Cellular Pharmacology Program, and ∥Human Proteomics Program, University of Wisconsin−Madison, Madison, Wisconsin 53719, United States
- Corresponding author
- Jin, Song
- Ge, Ying
- Venue
- Journal of the American Chemical Society
- Publisher
- American Chemical Society (ACS)
- Published
- 2015-02-11
- Volume
- 137
- Issue
- 7
- Pages
- 2432-2435
- ISSN
- 0002-7863 (ISSN-L)
- 1520-5126
- PMC
- PMC4372338
- OpenAlex
- W2326828010
- Cited by
- 70
- References
- 36
- Citations by year
- 2026: 4 · 2025: 1 · 2024: 6 · 2023: 4 · 2022: 6 · 2021: 3 · 2020: 9 · 2019: 6 · 2018: 8 · 2017: 10 · 2016: 9 · 2015: 4
- FWCI
- 4.60
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Journal of the American Chemical Society (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Heart, Lung, and Blood Institute (R01 HL096971, R01 HL109810)
- National Institute of Biomedical Imaging and Bioengineering (R21 EB013847)
- Topics
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Advanced Biosensing Techniques and Applications
- Physical Sciences › Chemistry › Analytical Chemistry › Analytical chemistry methods development
- Keywords
- phosphoprotein enrichment, protein phosphorylation, superparamagnetic nanoparticles, multivalent ligands, phosphoproteome, sample preparation
- MeSH terms
- Ligands, Phosphoproteins, Magnetite Nanoparticles
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