Identification of Post-Translational Modifications from Serum/Plasma by Immunoaffinity Enrichment and LC-MS/MS Analysis Without Depletion of Abundant Proteins

Hongbo Gu; Jianmin Ren; Xiaoying Jia; Matthew P Stokes

doi:10.1007/978-1-4939-7057-5_9

Identification of Post-Translational Modifications from Serum/Plasma by Immunoaffinity Enrichment and LC-MS/MS Analysis Without Depletion of Abundant Proteins

Methods Mol Biol. 2017:1619:119-125. doi: 10.1007/978-1-4939-7057-5_9.

Authors

Hongbo Gu¹, Jianmin Ren², Xiaoying Jia², Matthew P Stokes²

Affiliations

¹ Proteomic Service Group, Cell Signaling Technology, 3 Trask Lane, Danvers, MA, 01923, USA. Hongbo.gu@cellsignal.com.
² Proteomic Service Group, Cell Signaling Technology, 3 Trask Lane, Danvers, MA, 01923, USA.

PMID: 28674881
DOI: 10.1007/978-1-4939-7057-5_9

Abstract

Immunoaffinity enrichment combined with LC-MS/MS enables identification of Post-translational modifications (PTMs) from serum/plasma samples without abundant protein depletion. Here we described the workflow in details in identifying various types of PTMs such as lysine acetylation and arginine methylation from cancer serum. The method described is compatible with all common proteomic analysis platforms and quantitative methods.

Keywords: Immunoaffinity purification (IAP); LC-MS/MS; Post-translational modification (PTM); Serum/plasma.

MeSH terms

Blood Proteins* / chemistry
Blood Proteins* / isolation & purification
Chromatography, Liquid*
Humans
Protein Processing, Post-Translational*
Proteome*
Proteomics* / methods
Tandem Mass Spectrometry*

Substances

Blood Proteins
Proteome