Fig. 2.

Real time SPR pMHC stability assay. We immobilized each wildtype and heteroclitic pMHCI pair to the surface of a CM5 chip using streptavidin-biotin coupling to assess the half-life of each complex in real time. In a similar trend to the T2-binding assay, we observed that; (A) the natural tumor antigen, HLA A*0201-KIFGSLAFL, had a longer half-life compared to the heteroclitic variant, (A) HLA A*0201-KLFGSLAFV, whereas the natural tumor antigens (B) HLA A*0201-SLLMWITQC, (C) HLA A*0201-VISNDVCAQV and (D) HLA A*0201-YLEPGPVA had faster off-rates compared to their heteroclitic counterparts, (B) HLA A*0201-SLLMWITQL, (C) HLA A*0201-VLSNDVCAQV and (D) HLA A*0201-YLEPGPVV. Data are representative of 3 separate experiments.