Table 2.

Predicted and experimental peptide binding to HLA-A*0201

Peptide	BIMASa	SYFPEITHIa	MFI ratio Δ (n = 5)b	RA (n = 4)c	Stability DC50 (h)d
GVLVGVALI CEA694–702WT	8	23	0.92 ± 0.37	23.74 ± 14.12	<1
G L LVGVALI CEA694–702L2	89	29	1.18 ± 0.39	3.15 ± 3.82	1–2
GLLVGVAL V CEA694–702V9	52	25	1.87 ± 0.39	2.23 ± 0.89	2–4
G L LVGVAL V CEA694–702L2V9	592	31	1.78 ± 0.19	0.98 ± 0.17	4–6
GVL AR VALI CEA694AR4	8	21	n.d.	n.d.	n.d.
GVL AGM ALI CEA694AGM4	8	19	n.d.	n.d.	n.d.
GV VAL VALI CEA694VAL3	8	20	n.d.	n.d.	n.d.
IVGAETFYV HIVpol589–597	330	18	3.28 ± 0.61	–	n.d.

^aEach peptide sequence was entered into the BioInformatics and Molecular Analysis Section (BIMAS [26]) or the SYFPEITHI [30] algorithms. The HLA-A2 binding score output is annotated in the corresponding columns

^bValues are the ratio between anti-HLA-A2 mean fluorescence intensity measured on T2-cells loaded with saturating concentrations of peptide in serum-free media and non-loaded T2-cells

^cRA-relative HLA-A2 affinity values, which are calculated as described in the Materials and methods section

^dStability DC50 is the number of hours required to attain half maximal anti-HLA-A2 fluorescence at the surface of T2-cells at 37°C after removal of peptide from media