Related Experiment Video
Updated: Jul 6, 2025
07:35
Laser Microdissection-Based Protocol for the LC-MS/MS Analysis of the Proteomic Profile of Neuromelanin Granules
Published on: December 16, 2021
2.3K
Characterization of antigen presentation capability for neoantigen-based products using targeted LC-MS/MS method
Nan Xiang1, Kehua Zhang2, Yinghua Zhao3
1College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100049, China; The Cell Collection and Research Center, National Institutes for Food and Drug Control, No. 31 Huatuo St., Daxing District, Beijing, China.
Journal of Pharmaceutical and Biomedical Analysis
|January 7, 2024
Summary
We developed a sensitive mass spectrometry method to directly assess antigen presentation in neoantigen products. This technique enhances quality control by overcoming sensitivity limitations of previous methods.
Area of Science:
- Immunology
- Analytical Chemistry
- Biotechnology
Background:
- Neoantigen-based products require robust immune response assessment.
- Antigen presentation by antigen-presenting cells (APCs) is critical for immune response.
- Current bioassays for T-cell function have limitations.
- Mass spectrometry (MS) offers potential advantages for analyzing APCs but faces sensitivity challenges.
Purpose of the Study:
- To develop and validate a highly sensitive targeted liquid chromatography-tandem mass spectrometry (LC-MS/MS) method.
- To enable direct characterization of antigen presentation for quality control of neoantigen-based products.
- To overcome sensitivity limitations in MS-based analysis of APCs.
Main Methods:
- Optimized peptide extraction parameters for short peptides.
- Employed highly sensitive targeted triple quadrupole MS with ultra-high performance liquid chromatography (UHPLC).
- Utilized selective ion monitoring (Multiple Reaction Monitoring, MRM) and implemented quality control peptides.
- Validated the method according to ICH Q2 (R2) and M10 guidelines.
Main Results:
- Developed and validated a sensitive LC-MS/MS method for antigen presentation characterization.
- Demonstrated robustness and reliability using quality control peptides across different APCs.
- Achieved satisfactory validation results for selectivity, specificity, sensitivity, LOD, recovery, matrix effect, and repeatability.
- Successfully applied the method to dendritic cells (DCs) in the context of neoantigen-based products.
Conclusions:
- The developed targeted LC-MS/MS method offers a sensitive and robust approach for quality control of neoantigen-based products.
- This method directly assesses APCs' antigen-presenting capability, complementing traditional T-cell assays.
- Integration with T-cell assays provides a comprehensive evaluation of cell product quality from both physicochemical and biological aspects.
