MAGE-A4 TCR-like antibody immunoprecipitation in A375 and A375 MAGE-A4 KO xenograft tissue
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ABSTRACT: To demonstrate the on-target binding of the MAGE-A4 antibody to the target HLAp GVYDGREHTV, we developed a reverse immunopeptidomics strategy that uses the HLAp-targeting antibody as a bait to enrich interacting HLAp from A375 xenografts. For this purpose, we immobilized the IgG-format of the MAGE-A4 TCR-like antibody on Protein A beads by cross-linking, and exposed the molecule to the solubilized proteome, including membrane bound HLAp, of the A375 xenografts. We subsequently eluted the interacting HLAp bound to the MAGE-A4 antibody and purified the HLA-associated peptides through a 5 kDa molecular weight filter before liquid chromatography tandem mass spectrometry analysis for sequence determination.
INSTRUMENT(S): Q Exactive HF-X
ORGANISM(S): Homo Sapiens (human)
TISSUE(S): Skin
SUBMITTER: Nicola Ternette
LAB HEAD: Nicola Ternette
PROVIDER: PXD048297 | Pride | 2024-03-30
REPOSITORIES: Pride
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