A rapid method for cross-species quantitation of apolipoproteins A1, B48 and B100 in plasma by ultra-performance liquid chromatography/tandem mass spectrometry
Abstract
Apolipoprotein B100 (apoB100) and apolipoprotein A1 (apoA1) are the primary protein components of low density lipoprotein (LDL) and high density lipoprotein (HDL) particles, respectively, and plasma levels of these proteins are associated with risks of cardiovascular disease. Existing apoB100 quantitation methods for animal models have been limited to affinity capture techniques such as enzyme-linked immunosorbent assay (ELISA) and Western blot which require specialized reagents for each species and in many cases are not readily available. Here we demonstrate a single translatable ultra-performance liquid chromatography/tandem mass spectrometry (UPLC/MS/MS) assay that is fast and robust and can be used to measure apolipoprotein concentrations in plasma for six species. When possible, peptide sequences that are conserved across species were identified for this assay. The sample preparation is limited and can be carried out in 96-well microtiter plates and thus allows for multiplexed preparation of samples for analysis of large numbers of samples in a short time frame when combined with UPLC/MS/MS. Separation and quantitation of the tryptic peptides is carried out at 700 μL/min using a 1.7 µm core shell C18 column (2.1 × 50 mm). The chromatography is designed for the analysis of over 100 samples per day, and the UPLC run is less than 10 min. This assay is capable of supporting cardiovascular research by providing a single assay to measure critical biomarkers across multiple species without the need for antibodies, and does so in a high-throughput manner.
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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 [21]
Impairments of the BBB have been observed in the context of neurodegenerative diseases.
“impairments of the BBB have been observed in this context (Dickstein et al., 2006; Zipser et al., 2007; Lassman et al., 2012; Figure 2).”
From The impact of microglial activation on blood-brain barrier in brain diseases (2014) · cites this paper as Lassman et al., 2012
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- Authors
- Lassman, M. E., McLaughlin, T. M., Somers, E. P., Stefanni, A. C., Chen, Z., Murphy, B. A., Bierilo, K. K., Flattery, A. M., Wong, K. K., Castro‐Perez, J. M., Hubbard, B. K., & Roddy, T. P.
- Affiliations
- Merck & Co., Inc., Rahway, NJ, USA (United States) (US)
- Corresponding author
- Lassman, Michael E.
- Venue
- Rapid Communications in Mass Spectrometry
- Publisher
- Wiley
- Published
- 2011-12-05
- Volume
- 26
- Issue
- 2
- Pages
- 101-108
- ISSN
- 0951-4198 (ISSN-L)
- 1097-0231
- OpenAlex
- W1489764451
- Cited by
- 51
- References
- 24
- Citations by year
- 2023: 1 · 2022: 3 · 2021: 2 · 2020: 2 · 2018: 1 · 2017: 5 · 2016: 6 · 2015: 8 · 2014: 10 · 2013: 8 · 2012: 5
- FWCI
- 3.33
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Rapid Communications in Mass Spectrometry (publishedVersion)
- PubMed (publishedVersion)
- 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 › Metabolomics and Mass Spectrometry Studies
- Keywords
- apolipoprotein A-I, apolipoprotein B-48, apolipoprotein B-100, UPLC-MS/MS, cardiovascular biomarkers, low-density lipoprotein, high-density lipoprotein
- MeSH terms
- Amino Acid Sequence, Animals, Cardiovascular Diseases, Chromatography, High Pressure Liquid, Computer Simulation, Dogs, Cricetinae, Humans, Macaca mulatta, Peptide Fragments, Species Specificity, Trypsin, Linear Models, Apolipoprotein A-I, RNA, Small Interfering, Mice, Rats, Apolipoprotein B-48, Apolipoprotein B-100, Tandem Mass Spectrometry, Gene Knockdown Techniques
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