An MRM-Based Multiplexed Quantification Assay for Human Adipokines and Apolipoproteins

Area of Science:

• Biochemistry
• Proteomics
• Biomarker Discovery

Background:

• Adipokines and apolipoproteins are crucial in obesity and related diseases.
• Quantitative assessment of these proteins is vital for understanding disease mechanisms.
• ELISAs have limitations; mass spectrometry offers multiplexing and antibody-independent detection.

Purpose of the Study:

• To establish a multiple reaction monitoring (MRM)-based mass spectrometry assay for quantifying adipokines and apolipoproteins in human serum.
• To assess the feasibility of this assay for large cohort studies and diverse biological matrices.
• To investigate differences in protein abundance between normal weight and obese individuals.

Main Methods:

• Developed and optimized an MRM-based mass spectrometry method for 9 adipokines and 10 apolipoproteins.
• Implemented serum protein depletion to enhance detectability of low-abundance proteins.
• Validated the assay's accuracy and precision (intra-day and inter-day imprecision <16.5%).

Main Results:

• Quantified 8 adipokines and 10 apolipoproteins in 50 human serum samples.
• Observed significant abundance differences in five adipokines and four apolipoproteins between normal weight and obese groups.
• Demonstrated assay applicability to pediatric serum and cell culture supernatants.

Conclusions:

• The developed multiplexed MRM assay accurately quantifies key adipokines and apolipoproteins in human serum.
• This method facilitates large-scale investigations into protein-phenotype relationships relevant to obesity.
• The assay holds potential for future disease prediction strategies.