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.
Highlights
The percentage of overweight and obese people has significantly increased over the last decades, reaching epidemic dimensions [1,2]
We established a multiplexed multiple reaction monitoring (MRM)-based mass spectrometry (MS) assay for the quantification of circulating adipokines and apolipoproteins in different biological matrices
Sample Preparation and Peptide Selection was used for the quantification of 9 different adipokines and 10 apolipoproteins in serum samples of Previously, workflow for the quantification of the adipokine adiponectin
Summary
The percentage of overweight and obese people has significantly increased over the last decades, reaching epidemic dimensions [1,2]. The World Health Organization (WHO) classifies the nutritional status of adults by the BMI (body mass index) with overweight defined as BMI of. 25–29.9 kg/m2 and obesity as BMI ≥ 30 kg/m2 [3]. Molecules 2020, 25, 775 of white adipose tissue [4,5]. Adipose tissue was believed to be only responsible for energy storage [5,6]. The discovery of leptin revealed that adipose tissue signals its energy status to the brain. Leptin regulates appetite and satiety and energy expenditure and is directly proportional to the amount of adipose tissue [1,6]
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