Determination of HLA-DR binding of MAGE-A3 synthetic peptides corresponding to the sequences predicted by TEPITOPE to form promiscuous epitopes
| . | Sequences . | Predicted binding alleles* . | HLA-DRβ1 alleles . | ||||||
|---|---|---|---|---|---|---|---|---|---|
| *0101 . | *1501 . | *0301 . | *0401 . | *1101 . | *0701 . | *0801 . | |||
| Pool I residues | |||||||||
| 141-155 | GNWQYFFPVIFSKAS | 68 | 25 | 3 | >100‡ | 7 | 0.6 | 0.1 | 3.2 |
| 146-160 | FFPVIFSKASSSLQL | 66 | 10 | 0.24 | 7 | 2 | 1.8 | 0.01 | 1.5 |
| 156-170 | SSLQLVFGIELMEVD | 68 | 7 | 0.18 | 90 | 45 | 28 | 0.03 | 7 |
| 171-185 | PIGHLYIFATCLGLS | 96 | 0.3 | 0.028 | 2.8 | 0.9 | 0.9 | 0.01 | 1.5 |
| 281-295 | TSYVKVLHHMVKISG | 80 | 15 | 0.48 | 26 | 70 | 0.035 | 0.2 | 0.01 |
| Pool II residues | |||||||||
| 21-35 | EALGLVGAQAPATEE | 16 | 14 | 22 | >100‡ | >100‡ | >100‡ | 25 | >100‡ |
| 111-125 | RKVAELVHFLLLKYR | 76 | >100‡ | 0.055 | >100‡ | >100‡ | 0.7 | 55 | 7 |
| 161-175 | VFGIELMEVDPIGHL | 76 | >100‡ | 100 | 0.6 | 28 | >100‡ | 10 | 100 |
| 191-205 | GDNQIMPKAGLLIIV | 40 | >100‡ | 0.07 | >100‡ | >100‡ | 4 | 6 | 1 |
| 251-265 | VQENYLEYRQVPGSD | 48 | >100‡ | 5 | >100‡ | >100‡ | 60 | 26 | 10 |
| 286-300 | VLHHMVKISGGPHIS | 88 | 15 | 0.48 | >100‡ | >100‡ | 0.2 | 0.01 | 14 |
| HA (307-319)† | PKYVKQNTLKLAT | 84 | 0.18 | 2.4 | 6 | 0.9 | 0.7 | 0.18 | 3.2 |
| . | Sequences . | Predicted binding alleles* . | HLA-DRβ1 alleles . | ||||||
|---|---|---|---|---|---|---|---|---|---|
| *0101 . | *1501 . | *0301 . | *0401 . | *1101 . | *0701 . | *0801 . | |||
| Pool I residues | |||||||||
| 141-155 | GNWQYFFPVIFSKAS | 68 | 25 | 3 | >100‡ | 7 | 0.6 | 0.1 | 3.2 |
| 146-160 | FFPVIFSKASSSLQL | 66 | 10 | 0.24 | 7 | 2 | 1.8 | 0.01 | 1.5 |
| 156-170 | SSLQLVFGIELMEVD | 68 | 7 | 0.18 | 90 | 45 | 28 | 0.03 | 7 |
| 171-185 | PIGHLYIFATCLGLS | 96 | 0.3 | 0.028 | 2.8 | 0.9 | 0.9 | 0.01 | 1.5 |
| 281-295 | TSYVKVLHHMVKISG | 80 | 15 | 0.48 | 26 | 70 | 0.035 | 0.2 | 0.01 |
| Pool II residues | |||||||||
| 21-35 | EALGLVGAQAPATEE | 16 | 14 | 22 | >100‡ | >100‡ | >100‡ | 25 | >100‡ |
| 111-125 | RKVAELVHFLLLKYR | 76 | >100‡ | 0.055 | >100‡ | >100‡ | 0.7 | 55 | 7 |
| 161-175 | VFGIELMEVDPIGHL | 76 | >100‡ | 100 | 0.6 | 28 | >100‡ | 10 | 100 |
| 191-205 | GDNQIMPKAGLLIIV | 40 | >100‡ | 0.07 | >100‡ | >100‡ | 4 | 6 | 1 |
| 251-265 | VQENYLEYRQVPGSD | 48 | >100‡ | 5 | >100‡ | >100‡ | 60 | 26 | 10 |
| 286-300 | VLHHMVKISGGPHIS | 88 | 15 | 0.48 | >100‡ | >100‡ | 0.2 | 0.01 | 14 |
| HA (307-319)† | PKYVKQNTLKLAT | 84 | 0.18 | 2.4 | 6 | 0.9 | 0.7 | 0.18 | 3.2 |
The binding data are expressed in terms of relative binding capacity (IC50 μM), calculated as concentration of competitor peptide required to inhibit 50% of the binding of an allele specific biotinylated peptide (indicator peptide).
Percent of alleles predicted to bind the indicated sequences, with a threshold set at 5% (100% is represented by the 25 alleles incorporated in the software). The threshold is defined as the percentage of best scoring natural peptides and comprises a range between 1% to 10%; see “Materials and methods” for details.
HA307-319 is promiscuous sequence from influenza hemagglutinin.
IC50 values higher than 100 μM are outside the sensitivity limits of the binding assay.