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Peptide Identification Using Tandem Mass Spectrometry

Peptide Identification Using Tandem Mass Spectrometry

Tandem mass spectrometry, also known as MS/MS or MS2, is an analytical technique that employs two mass analyzers. Essentially it is a series of mass spectrometers that helps isolate a particular biomolecule and then helps study its chemical properties.
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...

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Updated: Feb 24, 2026

Phosphopeptide Enrichment Coupled with Label-free Quantitative Mass Spectrometry to Investigate the Phosphoproteome in Prostate Cancer

Phosphopeptide Enrichment Coupled with Label-free Quantitative Mass Spectrometry to Investigate the Phosphoproteome in Prostate Cancer

Published on: August 2, 2018

Sensitive, Robust, and Cost-Effective Approach for Tyrosine Phosphoproteome Analysis.

Mingming Dong^1,2, Yangyang Bian^1,3, Yan Wang^1,2

¹CAS Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences (CAS) , Dalian 116023, China.

Analytical Chemistry

|August 11, 2017

Summary

A new SH2 superbinder method enhances tyrosine phosphoproteome analysis, identifying 41% more phosphotyrosine (pTyr) peptides. This sensitive and cost-effective approach improves the study of cellular signaling pathways.

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Phosphopeptide Enrichment Coupled with Label-free Quantitative Mass Spectrometry to Investigate the Phosphoproteome in Prostate Cancer

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Published on: August 2, 2018

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Assessment of Resistance to Tyrosine Kinase Inhibitors by an Interrogation of Signal Transduction Pathways by Antibody Arrays

Assessment of Resistance to Tyrosine Kinase Inhibitors by an Interrogation of Signal Transduction Pathways by Antibody Arrays

Published on: September 19, 2018

Area of Science:

Biochemistry
Proteomics
Cellular Signaling

Background:

Tyrosine phosphorylation (pTyr) is crucial for cell signaling but challenging to analyze at the proteome level.
Conventional antibody-based methods for pTyr enrichment lack sensitivity, reproducibility, and are costly.

Purpose of the Study:

To develop and validate an efficient workflow for sensitive tyrosine phosphoproteome analysis using an SH2 domain-derived superbinder.
To overcome the limitations of traditional antibody-based enrichment methods for pTyr peptides.

Main Methods:

Utilized an SH2 superbinder for specific enrichment of phosphotyrosine (pTyr) peptides.
Applied the new workflow to analyze tyrosine phosphoproteome in various biological samples, including cell lines and tissue.

Main Results:

Identified 41% more pTyr peptides compared to previous methods.
Detected over 1800 high-confidence pTyr sites from 2 mg of pervanadate-treated Jurkat T cells.
Identified 343 high-confidence pTyr sites from 5 mg of unstimulated Jurkat cells, a 31% increase over antibody methods.
Characterized 197 high-confidence pTyr sites from mouse skeletal muscle tissue.

Conclusions:

The SH2 superbinder-based workflow offers a sensitive, robust, and cost-effective approach for tyrosine phosphoproteome analysis.
This method significantly enhances the identification of pTyr sites, even in low-abundance samples.
The approach has broad applications for studying the role of tyrosine phosphorylation in health and disease.