Fig 2. Epitope specificity.

The target HPV-E7p11-19 peptide sequence or the same sequence substituted with alanine at positions 1, 2, 3, 4, 5, 6, 7, 8, 9 indicated as A1 to A9, respectively, and pulsed onto T2 cells at 20ug/ml. The HPV sequence with the 4 middle amino acids substituted with alanine [HPV-E7AAAA] as used to gage binding to the center of the sequence as well. The binding of the mAbs 3F8 [A], 1B1 [B], or 2A5 [C] at a concentration of 3ug/ml was determined by flow cytometric analysis. T2 cells alone, or pulsed with RMF irrelevant peptide were the negative controls. The same cells were simultaneously stained with anti-HLA-A2 mAb, clone BB7.2, to measure the relative binding of the peptides to HLA-A2 molecule [D]. The data represent one of two similar experiments. [E]. Mirror plot of synthetic [bottom] and cell-derived experimental [top] YMLDLQPET peptides. Relative abundance refers to peak areas which are normalized to maximum peak.