Pre-fractionation Extends but also Creates a Bias in the Detectable HLA Class ? Ligandome.

ABSTRACT: HLA class ? molecules can communicate a range of cellular alterations (mutations, changes in protein copy number, aberrant post-translational modifications, or pathogen proteins) to CD8+ T lymphocytes in the form of HLA peptide ligands. At any given moment, tens of thousands of different self and foreign HLA class ? peptides may be presented on the cell surface by HLA class ? complexes. Due to the enormous biochemical diversity and low abundance of each of these peptides, HLA ligandome analysis presents unique challenges. Even with advances in enrichment strategies and MS instrumentation and fragmentation, sufficient ligandome depth for identification of viral pathogens and immuno therapeutically important tumor neo-antigens is still not routinely achievable. In this study, we evaluated two pre-fractionation techniques, high-pH reversed-phase and strong cation exchange, for the complementary analyses of HLA class ? peptide ligands. We observe that pre-fractionation substantially extends the detectable HLA class ? ligandome but also creates an identification bias. We thus advocate a rational choice between high-pH reversed-phase or strong cation exchange pre-fractionation for deeper HLA class ? ligandome analysis, depending on the HLA locus, allele, or peptide ligand modification in question.

SUBMITTER: Demmers LC 

PROVIDER: S-EPMC6456874 | biostudies-literature | 2019 Apr

REPOSITORIES: biostudies-literature