Related Experiment Video
Updated: May 12, 2025
09:32
Immunopeptidomics: Isolation of Mouse and Human MHC Class I- and II-Associated Peptides for Mass Spectrometry Analysis
Published on: October 15, 2021
11.3K
SAPrIm 2.0: a semi-automated protocol for mid-throughput soluble HLA immunopeptidomics
Erwin Tanuwidjaya1,2,3, Terry C C Lim Kam Sian1,2,3, Joel R Steele2
1Department of Medicine, School of Clinical Science, Faculty of Medicine, Nursing & Health Science, Monash University, Clayton, VIC, Australia.
Frontiers in Immunology
|May 9, 2025
Summary
We developed SAPrIm, an improved immunopeptidomics tool for analyzing soluble Human Leukocyte Antigen (HLA) molecules in plasma. This method offers reproducible profiling for cancer immunotherapy research.
Area of Science:
- Immunology
- Proteomics
- Cancer Research
Background:
- Human Leukocyte Antigen (HLA) molecules present peptides, forming the immunopeptidome, crucial for adaptive immunity and cancer immunotherapy.
- Plasma immunopeptidome profiling offers a minimally invasive method for monitoring disease and immune responses to cancer therapy.
Purpose of the Study:
- To introduce SAPrIm, an enhanced immunopeptidomics tool for soluble HLA analysis.
- To optimize the tool for high-throughput plasma immunopeptidome profiling.
Main Methods:
- The SAPrIm workflow was refined for soluble HLA analysis in plasma.
- The tool was validated for processing up to 12 samples per batch daily.
- Analysis of immunopeptides from 100 µL to 1 mL of plasma.
Main Results:
- Identified 1,200 to 4,000 immunopeptides per sample.
- Demonstrated high reproducibility across technical, biological, and inter-day analyses.
- Established strong potential for relative quantification in plasma-based studies.
Conclusions:
- SAPrIm provides a robust and reproducible method for plasma immunopeptidomics.
- The workflow facilitates efficient comparative analyses and mid-size cohort studies.
- This advancement supports the development of novel cancer immunotherapies.
