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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: Mar 30, 2026

Peptide:MHC Tetramer-based Enrichment of Epitope-specific T cells

Peptide:MHC Tetramer-based Enrichment of Epitope-specific T cells

Published on: October 22, 2012

Multiplexed Peptide-MHC Tetramer Staining with Mass Cytometry.

Mei Ling Leong¹, Evan W Newell²

¹School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, Singapore, Singapore, 637551.

Methods in Molecular Biology (Clifton, N.J.)

|November 7, 2015

Summary

Mass cytometry, a single-cell mass spectrometry technique, enables multiplexed assays. This study details methods for multiplexed peptide major histocompatibility (pMHC) tetramer staining using mass cytometry for antigen-specific cell detection.

Keywords:

Antigen-specific T cell Combinatorial tetramer staining Mass cytometry Peptide-MHC tetramer

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Area of Science:

Immunology
Analytical Chemistry
Biotechnology

Background:

Mass cytometry offers high-parameter single-cell analysis with reduced channel crosstalk.
Peptide major histocompatibility (pMHC) tetramer staining is crucial for identifying antigen-specific immune cells.
Multiplexing strategies are essential for comprehensive immune profiling.

Purpose of the Study:

To develop and describe methods for multiplexed pMHC-tetramer staining.
To leverage the capabilities of mass cytometry for advanced immune cell detection.
To enable combinatorial approaches for probing antigen-specific T cell populations.

Main Methods:

Utilized mass cytometry for high-dimensional single-cell analysis.
Applied peptide major histocompatibility (pMHC) tetramer staining techniques.
Developed and optimized multiplexing protocols for pMHC-tetramer staining in mass cytometry.

Main Results:

Successfully implemented multiplexed pMHC-tetramer staining using mass cytometry.
Demonstrated the feasibility of probing >40 parameters per cell.
Established a robust method for direct detection of antigen-specific cells with reduced crosstalk.

Conclusions:

Multiplexed pMHC-tetramer staining is effective with mass cytometry.
This method enhances the ability to identify and analyze antigen-specific immune cells.
The described techniques support combinatorial approaches for deeper immune system interrogation.