LARGE-SCALE PRODUCTION OF CLASS-I BOUND PEPTIDES - ASSIGNING A SIGNATURE TO HLA-B-ASTERISK-1501

A peptide-based vaccine must be bound and presented by major histocomp atibility complex class I molecules to elicit a CD8+ T-cell response. Because class I HLA molecules are highly polymorphic, it has yet to be established how well a vaccine peptide that stimulates one individual 's CD8+ cytotoxic T lymphocytes will be presented by a second individu al's different class I molecules. Therefore, to facilitate precise com parisons of class I peptide binding overlaps, we uniquely combined hol low-fiber bioreactors and mass spectrometry to assign precise peptide binding signatures to individual class I HLA molecules. In applying th is strategy to HLA-B1501, we isolated milligram quantities of B*1501- bound peptides and mapped them using mass spectrometry. Repeated analy ses consistently assign the same peptide binding signature to B1501; the degree of peptide binding overlap between any two class I molecule s can thus be determined through comparison of their peptide signature s.