The DOI matches its Crossref record; 33 of the 34 references it cites were found in a registry or an index.
Quantification of Thyroglobulin, a Low-Abundance Serum Protein, by Immunoaffinity Peptide Enrichment and Tandem Mass Spectrometry
Open accessAbstract
Background: Quantification of serum tumor markers plays an important role in determining whether patients treated for cancer require further therapy. Whereas large-scale proteomic efforts aim to identify novel tumor markers to facilitate early detection, optimization of methods for quantifying known tumor markers offers another approach to improving management of malignancies. For example, immunoassays used in clinical practice to measure established tumor markers suffer from potential interference from endogenous immunoglobulins and imperfect concordance across platforms—problems that also plague many other immunoassays. To address these important limitations, this study used peptide immunoaffinity enrichment in concert with liquid chromatography–tandem mass spectrometry (LC-MS/MS) to quantify thyroglobulin, a well-characterized tumor marker. Methods: We identified 3 peptides in tryptic digests of thyroglobulin that were detected at low concentrations by tandem mass spectrometry, raised polyclonal antibodies to those peptides, and used the antibodies to extract the 3 corresponding peptides from tryptic digests of human serum. We quantified each endogenous peptide using LC-MS/MS and multiple reaction monitoring with external calibrators. Results: The detection limit for endogenous thyroglobulin in serum was 2.6 μg/L (4 pmol/L). Direct comparison with immunoassay revealed good correlation (r2 = 0.81). Conclusions: Immunoaffinity peptide enrichment–tandem mass spectrometry can detect tryptic peptides of thyroglobulin at picomolar concentrations while also digesting the endogenous immunoglobulins that can potentially interfere with traditional immunoassays. Our observations suggest a general analytical strategy for using immunoaffinity isolation together with tandem mass spectrometry to quantify tumor antigens and other low-abundance proteins in human serum.
Overview
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Verification
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
In the citing paper
“Most known neosubstrate proteins have been identified through proteomic analyses of different cell lineages11,15,165,166.”
From Cancer therapies based on targeted protein degradation — lessons learned with lenalidomide (2021) · cites this paper as [166]
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Citations
Cites
Protein biomarker discovery and validation: the long and uncertain path to clinical utility
Rifai, N., Gillette, M. A., & Carr, S. A. (2006) · Nature Biotechnology · doi:10.1038/nbt1235 · Cited by 1,867 · cited 1×
“There is intense interest in using proteomics to discovernovelbiomarkersforthediagnosisandprognosis ofdisease;however,validationhasprovedtobeamajor stumblingblock( 1 ).Althoughthefinalaimofbiomarker discovery efforts is the development of a novel quantitative clinical assay, current immunoassay methodologies suffer fr…”
BackgroundLaboratory Support for the Diagnosis and Monitoring of Thyroid Disease
(2003) · Thyroid · doi:10.1089/105072503321086962 · Cited by 1,212
Anderson, N. L., Anderson, N. G., Haines, L. R., Hardie, D. B., Olafson, R. W., & Pearson, T. W. (2004) · Journal of Proteome Research · doi:10.1021/pr034086h · Cited by 806 · cited 2×
“We thereforetestedamethoddevelopedbyAndersonetal. ( 22 ), termed “stable isotope standards and capture by antipeptide antibodies,” that combines immunoaffinity peptide purification with tandem mass spectrometry.”
BackgroundKeshishian, H., Addona, T. A., Burgess, M. W., Kuhn, E. W., & Carr, S. A. (2007) · Molecular & Cellular Proteomics · doi:10.1074/mcp.m700354-mcp200 · Cited by 686 · cited 1×
“We therefore optimized the digestion of whole serum for the immunoaffinity enrichmentofTgpeptides.Oneimportantsourceofvariabilityinproteomicanalysesispartialproteolyticdigestion ofcomplexbiologicalmaterial.Highconcentrationsof chaotropic salts are commonly used to denature proteins and help facilitate more complete tr…”
MethodCrowther, J. R. (2000) · Humana Press eBooks · doi:10.1385/1592590497 · Cited by 517
Spencer, C. A., Takeuchi, M., Kazarosyan, M., Wang, C. C., Guttler, R., SINGER, P. A., Fatemi, S., LoPresti, J. S., & Nicoloff, J. T. (1998) · The Journal of Clinical Endocrinology & Metabolism · doi:10.1210/jcem.83.4.4683 · Cited by 476 · cited 1×
“Serum Tg has long been a thorn in the side of clinical tumor marker immunoassays because of its low concentration and common interferences (8, 10 , 11, 13, 26).”
BackgroundDemers, L. M., & Spencer, C. A. (2003) · Clinical Endocrinology · doi:10.1046/j.1365-2265.2003.01681.x · Cited by 460
Kuhn, E. W., Wu, J., Karl, J., Liao, H., Zolg, W., & Guild, B. (2004) · PROTEOMICS · doi:10.1002/pmic.200300670 · Cited by 402 · cited 2×
“To test a general approach to detecting low-abundance proteins in serum that avoids the limitations inherent to immunoassays, we took advantage of the power of mass spectrometry (MS) to detect specific peptidesincomplexmixtures.Previousattemptstouse MS to quantify low-abundance endogenous serum proteins include the di…”
BackgroundBarr, J. R., Maggio, V. L., Patterson, D. G., Cooper, G. R., Henderson, L. O., Turner, W. E., Smith, S. J., Hannon, W. H., Needham, L. L., & Sampson, E. J. (1996) · Clinical Chemistry · doi:10.1093/clinchem/42.10.1676 · Cited by 347
Kloos, R. T., & Mazzaferri, E. L. (2005) · The Journal of Clinical Endocrinology & Metabolism · doi:10.1210/jc.2005-0492 · Cited by 254
Research works printed but not linked: 4
- 1998, J Clin Endocrinol Metab, 83, 1121
- 1996, Ann Biol Clin (Paris), 54, 343
- 2006, Clin Lab Int, 8, 12
- 2001, The Elisa Guidebook, 421 p
Cited by
Sorted by citations
Haugen, B. R., Alexander, E. K., Bible, K. C., Doherty, G. M., Mandel, S. J., Nikiforov, Y. E., Pacini, F., Randolph, G. W., Sawka, A. M., Schlumberger, M., et al. (2016) · Thyroid® · doi:10.1089/thy.2015.0020 · Cited by 16,535
Selected reaction monitoring–based proteomics: workflows, potential, pitfalls and future directions
Picotti, P., & Aebersold, R. (2012) · Nature Methods · doi:10.1038/nmeth.2015 · Cited by 1,302
Addona, T. A., Abbatiello, S. E., Schilling, B., Skates, S. J., Mani, D. R., Bunk, D. M., Spiegelman, C. H., Zimmerman, L. J., Ham, A.-J. L., Keshishian, H., et al. (2009) · Nature Biotechnology · doi:10.1038/nbt.1546 · Cited by 1,037
The fundamental flaws of immunoassays and potential solutions using tandem mass spectrometry
Hoofnagle, A. N., & Wener, M. H. (2009) · Journal of Immunological Methods · doi:10.1016/j.jim.2009.06.003 · Cited by 562
“Incomplete digestion leads to variability in digestion from sample to sample, but despite this variability, multiple reports employing serum digestion and quantification by mass spectrometry have yielded promising results (Kuhn et al., 2004; Bondar et al., 2007; deWilde et al., 2008; Hoofnagle et al., 2008; Kuhn et al…”
BackgroundMethodInfluentialSelf-citationQuantitative analysis of peptides and proteins in biomedicine by targeted mass spectrometry
Gillette, M. A., & Carr, S. A. (2012) · Nature Methods · doi:10.1038/nmeth.2309 · Cited by 504
Metadata
- Authors
- Hoofnagle, A. N., Becker, J. O., Wener, M. H., & Heinecke, J. W.
- Affiliations
- Departments of Laboratory Medicine and
- University of Washington (US)
- Corresponding author
- Hoofnagle, Andrew N
- Venue
- Clinical Chemistry
- Publisher
- Oxford University Press (OUP)
- Published
- 2008-11-01
- Volume
- 54
- Issue
- 11
- Pages
- 1796-1804
- ISSN
- 0009-9147 (ISSN-L)
- 1530-8561
- PMC
- PMC2739673
- OpenAlex
- W2167033601
- Cited by
- 314
- References
- 34
- Citations by year
- 2026: 2 · 2025: 9 · 2024: 12 · 2023: 13 · 2022: 9 · 2021: 10 · 2020: 16 · 2019: 9 · 2018: 16 · 2017: 21 · 2016: 25 · 2015: 15 · 2014: 28 · 2013: 26 · 2012: 31
- FWCI
- 10.09
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model (vor)
- Locations
- Clinical Chemistry (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- University of Washington
- Topics
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Physical Sciences › Chemistry › Spectroscopy › Mass Spectrometry Techniques and Applications
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Advanced Biosensing Techniques and Applications
- Keywords
- thyroglobulin, tumor markers, immunoassay interference, polyclonal antibodies, tryptic peptides, multiple reaction monitoring, detection limit
- MeSH terms
- Amino Acid Sequence, Chromatography, Affinity, Chromatography, High Pressure Liquid, Humans, Immunoassay, Peptide Fragments, Peptide Mapping, Reference Standards, Sensitivity and Specificity, Thyroglobulin, Reproducibility of Results, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Tandem Mass Spectrometry
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
