Publication Details

Category Text Publication
Reference Category Journals
DOI 10.3390/molecules25040775
Licence creative commons licence
Title (Primary) An MRM-based multiplexed quantification assay for human adipokines and apolipoproteins
Author Krieg, L.; Schaffert, A.; Kern, M.; Landgraf, K.; Wabitsch, M.; Beck-Sickinger, A.G.; Koerner, A.; Blüher, M.; von Bergen, M.; Schubert, K.
Source Titel Molecules
Year 2020
Department MOLSYB
Volume 25
Issue 4
Page From art. 775
Language englisch
Keywords adipokines; apolipoproteins; multiple reaction monitoring; targeted LC-MS; obesity; human serum; SGBS cells; cell culture supernatant
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 interday 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.
Persistent UFZ Identifier
Krieg, L., Schaffert, A., Kern, M., Landgraf, K., Wabitsch, M., Beck-Sickinger, A.G., Koerner, A., Blüher, M., von Bergen, M., Schubert, K. (2020):
An MRM-based multiplexed quantification assay for human adipokines and apolipoproteins
Molecules 25 (4), art. 775 10.3390/molecules25040775