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Enrichment of Phosphorylated MHC Peptides with Immobilized Metal Affinity Chromatography and Titanium Dioxide
1Human Health Therapeutics Research Centre, National Research Council Canada, Ottawa, ON, Canada.
Abstract:
Phosphorylation is one of the most important forms of posttranslational modification. Dysregulation of phosphorylation is implicated in tumorigenesis, with cancerous signaling pathways activated by kinases. For immunotherapy with neoantigen-based peptides, phosphopeptides derived from aberrantly phosphorylated proteins presented by major histocompatibility complex (MHC) are promising candidates due to their specificity to elicit cytotoxic T-cell responses. Unlike other MHC peptides, phosphorylated MHC peptides cannot be predicted from DNA sequences, and their identification relies on the direct detection of phosphopeptides using mass spectrometry (MS). For MS detection, it is extremely important to enrich phosphorylated peptides from the complex repertoire of MHC peptides. Herein, we describe the combined use of immobilized metal affinity chromatography and titanium dioxide nanoparticles for phosphopeptides enrichment from immunopeptidome.
Insights
This study introduces a novel method for enriching phosphopeptides from the immunopeptidome. This technique enhances the identification of potential neoantigens for cancer immunotherapy by improving mass spectrometry detection.
Area of Science:
- Biochemistry
- Immunology
- Cancer Research
Background:
- Phosphorylation is a key posttranslational modification implicated in cancer signaling pathways.
- Aberrantly phosphorylated proteins presented by major histocompatibility complex (MHC) are promising targets for neoantigen-based immunotherapy.
- Identifying these phosphopeptides is challenging as they cannot be predicted from DNA sequences.
Purpose of the Study:
- To develop an effective method for enriching phosphopeptides from the immunopeptidome for mass spectrometry (MS) detection.
- To improve the identification of specific phosphopeptides for cancer immunotherapy.
Main Methods:
- Combined use of immobilized metal affinity chromatography (IMAC) and titanium dioxide (TiO2) nanoparticles.
- Enrichment of phosphopeptides from the complex immunopeptidome.
Main Results:
- Successfully enriched phosphopeptides from the immunopeptidome.
- The described method facilitates the direct detection of phosphopeptides using MS.
Conclusions:
- The combined IMAC and TiO2 nanoparticle approach is effective for phosphopeptide enrichment.
- This method aids in the identification of potential neoantigens for targeted cancer immunotherapy.
