Figure 1.

HLAII immunopeptidomics workflow for the identification of peptides presented from TNF antagonists by DCs. Freshly isolated PBMCs collected from fourteen healthy donors were isolated by density separation for subsequent CD14+ monocyte-purification. CD14+ monocytes were cultured and differentiated into DCs for six days. DCs were loaded with TNF antagonists or their preformed TNF immune complexes at day 5, followed by LPS stimulation for 22 h. DCs were harvested, washed and lysed to obtain whole protein extracts for HLAII immunoprecipitation using a pan-HLAII antibody. The HLAII immunopeptidome was purified by filtration and by C18 prior to LC-MS/MS analysis using trapped ion mobility time-of-flight mass spectrometry (timsTOF MS).