Isotope dilution--mass spectrometric quantification of specific proteins: model application with apolipoprotein A-I
Abstract
An enzymatic hydrolysis isotope dilution-mass spectrometric method was developed for reference quantification of specific proteins. The analytical procedure involved measuring a reproducibly hydrolyzed peptide (serving as the primary standard) unique to a specific protein. This new mass spectrometric method was evaluated by assessing the concentration of apolipoprotein (apo) A-I in the European Community Bureau of Reference (BCR) lyophilized Certified Reference Material (CRM 393). We used the method to make 96 measurements (4 replicate analyses of 4 enzymatic digests of 6 vials of BCR-CRM 393), which gave an average total protein mass of 1.048 mg (+/- 1.0% at 99% confidence limits). The total overall analytical CV was 3.95%. The results of this evaluation of our model approach to determine the concentration of a specific protein in a purified preparation demonstrated that our new mass spectrometric method can be used to measure apolipoproteins and other specific proteins without the use of epitopic immunoassay methods.
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Various LC-MS/MS assays for multiplexed quantification of apolipoproteins or phenotyping of apoE have been reported.
“Various LC-MS/MS assays for multiplexed quantification of apolipoproteins (14-23) or phenotyping of apoE (24-27) have been reported.”
From Automated multiplex LC-MS/MS assay for quantifying serum apolipoproteins A-I, B, C-I, C-II, C-III, and E with qualitative apolipoprotein E phenotyping (2016) · cites this paper as [23]
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- Authors
- Barr, 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.
- Affiliations
- Centers for Disease Control and Prevention (US)
- ORCID
- Barr, J R
- Venue
- Clinical Chemistry
- Publisher
- Oxford University Press (OUP)
- Published
- 1996-10-01
- Volume
- 42
- Issue
- 10
- Pages
- 1676-1682
- ISSN
- 0009-9147 (ISSN-L)
- 1530-8561
- OpenAlex
- W1892810170
- Cited by
- 347
- References
- 0
- Citations by year
- 2026: 4 · 2024: 1 · 2023: 2 · 2022: 4 · 2021: 5 · 2020: 4 · 2019: 4 · 2018: 1 · 2017: 16 · 2016: 19 · 2015: 20 · 2014: 29 · 2013: 26 · 2012: 34
- FWCI
- 1.22
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model (vor)
- Locations
- Clinical Chemistry (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Physical Sciences › Chemistry › Spectroscopy › Mass Spectrometry Techniques and Applications
- Social Sciences › Decision Sciences › Statistics, Probability and Uncertainty › Scientific Measurement and Uncertainty Evaluation
- Keywords
- isotope dilution mass spectrometry, apolipoprotein A-I, protein quantification, enzymatic hydrolysis, certified reference materials, apolipoproteins, reference measurement procedure
- MeSH terms
- Amino Acid Sequence, Humans, Hydrolysis, Indicator Dilution Techniques, Molecular Sequence Data, Molecular Weight, Peptide Fragments, Peptide Mapping, Mass Spectrometry, Trypsin, Spectrometry, Mass, Fast Atom Bombardment, Apolipoprotein A-I
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