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Variability in Mass Spectrometry-based Quantification of Clinically Relevant Drug Transporters and Drug Metabolizing Enzymes

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Author: Wegler, C. · Gaugaz, F.Z. · Andersson, T.B. · Wiśniewski, J.R. · Busch, D. · Gröer, C. · Oswald, S. · Norén, A. · Weiss, F. · Hammer, H.S. · Joos, T.O. · Poetz, O. · Achour, B. · Rostami-Hodjegan, A. · Steeg, E. van de · Wortelboer, H.M. · Artursson, P.
Type:article
Date:2017
Source:Molecular Pharmaceutics, 9, 14, 3142-3151
Identifier: 781416
doi: doi:10.1021/acs.molpharmaceut.7b00364
Keywords: Biology · Drug metabolizing enzymes · Drug transporters · Label-free proteomics · Membrane proteins · Protein quantification · Targeted proteomics · Biomedical Innovation · Healthy Living · Life · MSB - Microbiology and Systems Biology · ELSS - Earth, Life and Social Sciences

Abstract

Many different methods are used for mass-spectrometry-based protein quantification in pharmacokinetics and systems pharmacology. It has not been established to what extent the results from these various methods are comparable. Here, we compared six different mass spectrometry-based proteomics methods by measuring the expression of clinically relevant drug transporters and metabolizing enzymes in human liver. Mean protein concentrations were in general quantified to similar levels by methods using whole tissue lysates. Methods using subcellular membrane fractionation gave incomplete enrichment of the proteins. When the enriched proteins were adjusted to levels in whole tissue lysates, they were on average 4-fold lower than those quantified directly in whole tissue lysates. The differences in protein levels were propagated into differences in predictions of hepatic clearance. In conclusion, caution is needed when comparing and applying quantitative proteomics data obtained with different methods, especially since membrane fractionation is common practice for protein quantification used in drug clearance predictions. © 2017 American Chemical Society.