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Serum/Plasma Proteomics pp 417–430Cite as

Absolute Quantification of Middle- to High-Abundant Plasma Proteins via Targeted Proteomics

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1619))

Abstract

The increasing number of peptide and protein biomarker candidates requires expeditious and reliable quantification strategies. The utilization of liquid chromatography coupled to quadrupole tandem mass spectrometry (LC-MS/MS) for the absolute quantitation of plasma proteins and peptides facilitates the multiplexed verification of tens to hundreds of biomarkers from smallest sample quantities. Targeted proteomics assays derived from bottom-up proteomics principles rely on the identification and analysis of proteotypic peptides formed in an enzymatic digestion of the target protein. This protocol proposes a procedure for the establishment of a targeted absolute quantitation method for middle- to high-abundant plasma proteins waiving depletion or enrichment steps. Essential topics as proteotypic peptide identification and LC-MS/MS method development as well as sample preparation and calibration strategies are described in detail.

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Acknowledgment

This publication is supported by LIFE—Leipzig Research Center for Civilization Diseases, Universität Leipzig. LIFE is funded by means of the European Union, by the European Regional Development Fund (ERDF), and by the Free State of Saxony within the framework of the excellence initiative.

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Authors and Affiliations

  1. Institute of Laboratory Medicine, Clinical Chemistry and Molecular Diagnostics, University Hospital Leipzig, Paul-List-Str. 13-15, Leipzig, 04103, Germany

    Julia Dittrich & Uta Ceglarek

  2. LIFE-Leipzig Research Center for Civilization Diseases, Leipzig University, Philipp-Rosenthal-Str. 27, Leipzig, Germany

    Julia Dittrich & Uta Ceglarek

Authors
  1. Julia Dittrich
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  2. Uta Ceglarek
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Corresponding author

Correspondence to Julia Dittrich .

Editor information

Editors and Affiliations

  1. Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Melbourne, Victoria, Australia

    David W. Greening

  2. Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Melbourne, Victoria, Australia

    Richard J. Simpson

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Dittrich, J., Ceglarek, U. (2017). Absolute Quantification of Middle- to High-Abundant Plasma Proteins via Targeted Proteomics. In: Greening, D., Simpson, R. (eds) Serum/Plasma Proteomics. Methods in Molecular Biology, vol 1619. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7057-5_29

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  • DOI: https://doi.org/10.1007/978-1-4939-7057-5_29

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7056-8

  • Online ISBN: 978-1-4939-7057-5

  • eBook Packages: Springer Protocols

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