Isolation of HLA Bound Peptides by Immunoaffinity Capture and Identification by Mass Spectrometry

Kirti Pandey, Sri H. Ramarathinam, Anthony W. Purcell

Research output: Contribution to journalArticleOtherpeer-review

11 Citations (Scopus)


This article describes the purification of HLA-bound peptides and their subsequent sequencing by mass spectrometry. These methods can be used for both HLA class I and class II molecules and can be adapted to different species depending on the availability of specific antibodies. Peptides can be successfully isolated from a variety of sample types, including in vitro cultured cells and primary tissues. The method involves the affinity capture of HLA-peptide complexes and separation of peptides from HLA heavy chains, followed by tailored interrogation by mass spectrometry to take into account the non-tryptic nature of endogenously derived HLA-bound peptides.

Original languageEnglish
Article numbere92
Number of pages18
JournalCurrent Protocols
Issue number3
Publication statusPublished - Mar 2021


  • antigen presentation
  • HLA peptide
  • immunopeptidome
  • mass spectrometry

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