Abstract
Immunoaffinity enrichment of proteotypic peptides, coupled with selected reaction monitoring, enables indirect protein quantification. However the lack of suitable antibodies limits its widespread application. We developed a method in which multi-specific antibodies are used to enrich groups of peptides, thus facilitating multiplexed quantitative protein assays. We tested this strategy in a pharmacokinetic experiment by targeting a group of homologous drug transforming proteins in human hepatocytes. Our results indicate the generic applicability of this method to any biological system.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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ATP Binding Cassette Transporter, Subfamily B / immunology
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ATP Binding Cassette Transporter, Subfamily B / metabolism
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Amino Acid Sequence
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Antibodies / immunology
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Antibody Affinity
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Aryl Hydrocarbon Hydroxylases / immunology
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Aryl Hydrocarbon Hydroxylases / metabolism
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Atorvastatin / pharmacokinetics
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Cells, Cultured
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Chromatography, Liquid / methods
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Cytochrome P-450 CYP3A / immunology
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Cytochrome P-450 CYP3A / metabolism
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Epitopes / immunology
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Epitopes / metabolism
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Hepatocytes / cytology
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Hepatocytes / enzymology*
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Hepatocytes / metabolism*
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Humans
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Hydroxymethylglutaryl-CoA Reductase Inhibitors / pharmacokinetics
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Mass Spectrometry / methods*
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Peptides / immunology
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Peptides / metabolism*
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Pravastatin / pharmacokinetics
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Primary Cell Culture
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Sequence Homology, Amino Acid
Substances
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ABCB1 protein, human
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ATP Binding Cassette Transporter, Subfamily B
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Antibodies
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Epitopes
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Hydroxymethylglutaryl-CoA Reductase Inhibitors
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Peptides
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Atorvastatin
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Aryl Hydrocarbon Hydroxylases
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CYP3A7 protein, human
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Cytochrome P-450 CYP3A
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Pravastatin