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Peptide Identification Using Tandem Mass Spectrometry01:33

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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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Related Experiment Video

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Imaging the Human Immunological Synapse
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Tsyn-Seq: a T-cell Synapse-Based Antigen Identification Platform.

Yimei Jin1, Takahiko Miyama1, Alexandria Brown1

  • 1Department of Genomic Medicine, The University of Texas MD Anderson Cancer Center, Houston, Texas.

Cancer Immunology Research
|February 16, 2024
PubMed
Summary

We developed T-synapse sequencing (Tsyn-seq), a novel method for rapidly identifying T-cell receptor (TCR) target antigens. This system successfully identified HPV16 E7 antigen from a cervical cancer cell line, aiding cancer research.

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Area of Science:

  • Immunology
  • Molecular Biology
  • Genomics

Background:

  • Identifying T-cell receptor (TCR) targets is crucial for understanding immune responses.
  • Existing methods for genome-wide antigen identification are not universally accessible.

Purpose of the Study:

  • To present a new, rapid, and universally applicable method for identifying peptide-major histocompatibility complex (pMHC) targets recognized by TCRs.
  • To enable functional screening of target antigens from cDNA libraries.

Main Methods:

  • Developed the T-synapse (Tsyn) reporter system using antigen-presenting cells (APCs) with an NF-κB reporter and T cells with an NFAT reporter.
  • Enriched interacting T cell-APC aggregates via dual-reporter activity and flow sorting.
  • Quantified antigen identification using deep sequencing (Tsyn-seq).

Main Results:

  • Tsyn-seq successfully enriched the cognate antigen (HPV16 E7) for a previously characterized TCR.
  • The method was validated using a cDNA library from an HPV16-positive cervical cancer cell line.
  • Demonstrated rapid identification of TCR-recognized antigens.

Conclusions:

  • Tsyn-seq offers a powerful new tool for identifying TCR target antigens.
  • This method facilitates rapid screening of tumor cDNA libraries for relevant antigens.
  • Advances TCR-antigen discovery in cancer immunology and immunotherapy research.