Variability assessment of 15N metabolic labeling-based proteomics workflow in mouse plasma and brain

Abstract

¹⁵N metabolic labeling-based quantitative proteomics is used for the identification of disease- and phenotype-related alterations in live organisms. The variability of ¹⁵N metabolic labeling proteomics workflows has been assessed in plants and bacteria. However, no study has addressed this topic in mice. We have investigated the repeatability of a quantitative in vivo¹⁵N metabolic labeling proteomics workflow in mice by assessing LC variability, peptide and protein profiling characteristics and overall ¹⁵N/¹⁴N protein quantification accuracy in technical replicates of plasma and brain specimens. We furthermore examined how sample preparation affects these parameters in plasma and brain. We found that specimen type (i.e. plasma or brain) influences the variability of the ¹⁵N metabolic labeling workflow in an LC-independent manner.