Proteomics

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MAGE-A4 TCR-like antibody immunoprecipitation in A375 and A375 MAGE-A4 KO xenograft tissue


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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