An MRM-Based multiplexed quantification assay for human adipokines and apolipoproteins
peer-reviewed
Erstveröffentlichung
2020-02-11Authors
Krieg, Laura
Schaffert, Alexandra
Kern, Matthias
Landgraf, Kathrin
Wabitsch, Martin
Wissenschaftlicher Artikel
Published in
Molecules ; 25 (2020), 4. - Art.-Nr. 775. - eISSN 1420-3049
Link to original publication
https://dx.doi.org/10.3390/molecules25040775Institutions
UKU. Klinik für Kinder- und JugendmedizinExternal cooperations
Helmholtz-Zentrum Umweltforschung GmbH UFZUniversität Leipzig
Document version
published version (publisher's PDF)Abstract
Adipokines and apolipoproteins are key regulators and potential biomarkers in obesity and associated diseases and their quantitative assessment is crucial for functional analyses to understand disease mechanisms. Compared to routinely used ELISAs, multiple reaction monitoring (MRM)-based mass spectrometry allows multiplexing and detection of proteins for which antibodies are not available. Thus, we established an MRM method to quantify 9 adipokines and 10 apolipoproteins in human serum. We optimized sample preparation by depleting the two most abundant serum proteins for improved detectability of low abundant proteins. Intra-day and inter-day imprecision were below 16.5%, demonstrating a high accuracy. In 50 serum samples from participants with either normal weight or obesity, we quantified 8 adipokines and 10 apolipoproteins. Significantly different abundances were observed for five adipokines (adipsin, adiponectin, chemerin, leptin, vaspin) and four apolipoproteins (apo-B100/-C2/-C4/-D) between the body mass index (BMI) groups. Additionally, we applied our MRM assay to serum samples from normal weight children and human adipocyte cell culture supernatants to proof the feasibility for large cohort studies and distinct biological matrices. In summary, this multiplexed assay facilitated the investigation of relationships between adipokines or apolipoproteins and phenotypes or clinical parameters in large cohorts, which may contribute to disease prediction approaches in the future.
Subject headings
[GND]: Adipokine | Apolipoproteine | LC-MS | Fettsucht | Blutserum[MeSH]: Adipokines | Apolipoproteins | Chromatography, Liquid | Mass spectrometry | Obesity | Serum
[Free subject headings]: Multiple reaction monitoring | Targeted LC-MS | Human serum | SGBS cells | Cell culture supernatant
[DDC subject group]: DDC 570 / Life sciences | DDC 610 / Medicine & health
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Please use this identifier to cite or link to this item: http://dx.doi.org/10.18725/OPARU-39485
Krieg, Laura et al. (2021): An MRM-Based multiplexed quantification assay for human adipokines and apolipoproteins. Open Access Repositorium der Universität Ulm und Technischen Hochschule Ulm. http://dx.doi.org/10.18725/OPARU-39485
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