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  Quantitative Proteomics of Secreted Proteins.

Frauenstein, A., & Meissner, F. (2018). Quantitative Proteomics of Secreted Proteins. In D. De Nardo, & C. De Nardo (Eds.), Innate Immune Activation (pp. 215-227). New York, NY: Humana Press.

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 Creators:
Frauenstein, Annika1, Author              
Meissner, Felix1, Author              
Affiliations:
1Meissner, Felix / Experimental Systems Immunology, Max Planck Institute of Biochemistry, Max Planck Society, ou_2149678              

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Free keywords: Mass spectrometry Quantitative proteomics Label-free quantification Secretome Secreted proteins Cytokines Interleukins Interferons Growth factors Sample preparation
 Abstract: Secreted proteins such as cytokines, interleukins, growth factors, and hormones have pleiotropic functions and facilitate intercellular communication in organisms. Quantification of these proteins conventionally relies on antibody-based methods, i.e., enzyme-linked immunosorbent assays (ELISA), whose large-scale use is limited by availability, specificity, and affordability. Here, we describe an experimental and bioinformatics workflow to comprehensively quantify cellular protein secretion by mass spectrometry. Secreted proteins are collected in vitro or ex vivo, digested with proteases and the resulting peptide mixtures are analyzed in single liquid chromatography–mass spectrometry (LC-MS/MS) runs. Label-free quantification and bioinformatics analysis is conducted in the MaxQuant and Perseus computational environment. Our workflow allows the quantification of thousands of secreted proteins spanning a concentration range of four orders of magnitude and permits the systems-level characterization of secretory programs as well as the discovery of proteins with unexpected extracellular functions.

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 Dates: 2018
 Publication Status: Published in print
 Pages: -
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 Rev. Type: -
 Identifiers: ISI: 29177865
DOI: 10.1007/978-1-4939-7519-8_14
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Title: Innate Immune Activation
Source Genre: Book
 Creator(s):
De Nardo, D., Editor
De Nardo, C., Editor
Affiliations:
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Publ. Info: New York, NY : Humana Press
Pages: - Volume / Issue: - Sequence Number: - Start / End Page: 215 - 227 Identifier: -

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Title: Methods in Molecular Biology
Source Genre: Series
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Publ. Info: Humana Press
Pages: - Volume / Issue: 1714 Sequence Number: - Start / End Page: - Identifier: -