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Abstract:
N-15 metabolic labeling-based quantitative proteomics is used for the
identification of disease-and phenotype-related alterations in live
organisms. The variability of N-15 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 vivo N-15 metabolic labeling proteomics workflow in
mice by assessing LC variability, peptide and protein profiling
characteristics and overall N-15/N-14 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 N-15 metabolic labeling workflow in an LC-independent
manner.
