Protocol for hybrid mass spectrometry-AI mapping of proteasome-generated HLA class I epitopes
Proteasomes play a central role in antiviral and antitumor cytotoxic immunity, generating most of the >10,000 unique proteome-derived peptides bound to ∼200,000 human leukocyte antigen (HLA) class I molecules on each human cell. Here, we present a protocol for hybrid mass spectrometry-AI mapping of proteasome-generated HLA class I epitopes. We describe steps for in vitro processing of antigens by proteasomes, HLA class I allele selection, and peptide pool preparation. We then detail procedures for determining HLA class I ligands and population immunity. For complete details on the use and execution of this protocol, please refer to Kudriaeva et al.<sup>1</sup>.
Authors
- Maxim Ri (ORCID: https://orcid.org/0000-0003-2403-884X)
- Anna A. Kudriaeva (ORCID: https://orcid.org/0000-0003-1163-9929)
- Alexey A. Belogurov (ORCID: https://orcid.org/0000-0002-2033-9621)
- George A. Saratov (ORCID: https://orcid.org/0000-0003-1048-2405)
Institutions
- Institute of Bioorganic Chemistry (RU)
- Institut de Biologie Moléculaire et Cellulaire (FR)
Publication Details
- Journal
- STAR Protocols
- Published
- 2025-10-23
- DOI
- https://doi.org/10.1016/j.xpro.2025.104154
- Primary Topic
- vaccines and immunoinformatics approaches
- Type
- article
- Field-Weighted Citation Impact
- 0.00