Detection of oxidized and glycated proteins in clinical samples using mass spectrometry — A user's perspective
Abstract
Background Proteins in human tissues and body fluids continually undergo spontaneous oxidation and glycation reactions forming low levels of oxidation and glycation adduct residues. Proteolysis of oxidised and glycated proteins releases oxidised and glycated amino acids which, if they cannot be repaired, are excreted in urine. Scope of review In this review we give a brief background to the classification, formation and processing of oxidised and glycated proteins in the clinical setting. We then describe the application of stable isotopic dilution analysis liquid chromatography-tandem mass spectrometry (LC-MS/MS) for measurement of oxidative and glycation damage to proteins in clinical studies, sources of error in pre-analytic processing, corroboration with other techniques - including how this may be improved - and a systems approach to protein damage analysis for improved surety of analyte estimations. Major conclusions Stable isotopic dilution analysis LC-MS/MS provides a robust reference method for measurement of protein oxidation and glycation adducts. Optimised pre-analytic processing of samples and LC-MS/MS analysis procedures are required to achieve this. General significance Quantitative measurement of protein oxidation and glycation adducts provides information on level of exposure to potentially damaging protein modifications, protein inactivation in ageing and disease, metabolic control, protein turnover, renal function and other aspects of body function. Reliabl…
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Use of chromatography-based methods in routine clinical studies is limited, particularly due to high costs.
“but their use in routine clinical studies is limited, in particular, due to high costs [158,163,164]”
From Protein oxidation - Formation mechanisms, detection and relevance as biomarkers in human diseases (2021) · cites this paper as [163]
Chromatography-based methods (HPLC, UPLC, mainly combined with MS/MS) are the most sensitive approaches for detection of glycoxidation products.
“The most sensitive approaches for detection of glycoxidation products are methods based on chromatography, including HPLC und UPLC that are mainly combined with MS/MS.”
From Protein oxidation - Formation mechanisms, detection and relevance as biomarkers in human diseases (2021) · cites this paper as [163]
Chromatography-based methods represent the gold standard for identification of glycoxidation structures and detection in free and protein-bound forms.
“These methods represent the gold standard for the identification of glycoxidation structures and detection in free and protein-bound forms”
From Protein oxidation - Formation mechanisms, detection and relevance as biomarkers in human diseases (2021) · cites this paper as [163]
Cites
Pérez, V. I., Buffenstein, R., Masamsetti, V., Leonard, S., Salmon, A. B., Mele, J., Andziak, B., Yang, T., Edrey, Y., Friguet, B., et al. (2009) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0809620106 · Cited by 418 · see citing works
Glycation free adduct accumulation in renal disease: the new AGE
Thornalley, P. J. (2005) · Pediatric Nephrology · doi:10.1007/s00467-005-2011-9 · Cited by 53 · see citing works
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Advanced glycation end products in the pathogenesis of chronic kidney disease
Rabbani, N., & Thornalley, P. J. (2018) · Kidney International · doi:10.1016/j.kint.2017.11.034 · Cited by 387 · see citing works
Self-citationProtein oxidation - Formation mechanisms, detection and relevance as biomarkers in human diseases
Kehm, R., Baldensperger, T., Raupbach, J., & Höhn, A. (2021) · Redox Biology · doi:10.1016/j.redox.2021.101901 · Cited by 348 · see citing works
“These methods represent the gold standard for the identification of glycoxidation structures and detection in free and protein-bound forms, but their use in routine clinical studies is limited, in particular, due to high costs [158,163,164].”
MethodRicharme, G., Mihoub, M., Dairou, J., Bui, L. C., Leger, T., & Lamouri, A. (2015) · Journal of Biological Chemistry · doi:10.1074/jbc.m114.597815 · Cited by 268 · see citing works
Verrastro, I., Pasha, S., Jensen, K., Pitt, A., & Spickett, C. (2015) · Biomolecules · doi:10.3390/biom5020378 · Cited by 114 · see citing works
Ashraf, J. M., Ahmad, S., Choi, I., Ahmad, N., Farhan, M., Tatyana, G., & Shahab, U. (2015) · IUBMB Life · doi:10.1002/iub.1450 · Cited by 106 · see citing works
Teichert, T., Hellwig, A., Peßler, A., Hellwig, M., Vossoughi, M., Sugiri, D., Vierkötter, A., Schulte, T., Freund, J., Roden, M., et al. (2015) · PLOS ONE · doi:10.1371/journal.pone.0128293 · Cited by 20 · see citing works
- Authors
- Thornalley, P. J., & Rabbani, N.
- Affiliations
- University Hospital Coventry (GB)
- Corresponding author
- Thornalley, Paul J.
- Venue
- Biochimica et Biophysica Acta (BBA) - General Subjects
- Publisher
- Elsevier BV
- Published
- 2014-02
- Volume
- 1840
- Issue
- 2
- Pages
- 818-829
- ISSN
- 0304-4165 (ISSN-L)
- OpenAlex
- W2107524932
- Cited by
- 133
- References
- 2
- Citations by year
- 2026: 1 · 2025: 3 · 2024: 9 · 2023: 14 · 2022: 10 · 2021: 8 · 2020: 8 · 2019: 10 · 2018: 13 · 2017: 11 · 2016: 17 · 2015: 18 · 2014: 8 · 2013: 2
- FWCI
- 1.56
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Biochimica et Biophysica Acta (BBA) - General Subjects (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Clinical Biochemistry › Advanced Glycation End Products research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Keywords
- protein oxidation, protein glycation, LC-MS/MS, clinical diagnostics, metabolic control, renal function, protein turnover, immunoassay comparison
- MeSH terms
- Humans, Oxidation-Reduction, Proteins, Mass Spectrometry, Glycation End Products, Advanced
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