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. 2022 Mar 17;17(3):e0265534. doi: 10.1371/journal.pone.0265534

Fig 1. Binding of the mAbs to the HPV-E7p11-19/HLA-A2 complex.

Fig 1

[A] Binding of mAbs to T2 cells pulsed with or without peptides. HPV-E7p11, HPV-E7AAAA or, WT1-RMF peptide at a concentration of 20ug/ml was pulsed onto T2 cells overnight in serum-free RPMI1640 complete medium. Cells were washed and stained with the mAbs 1B1, 2A5 or 3F8 conjugated to APC at a concentration of 3ug/ml. A control TCRm mAb specific for WT1-RMF/HLA-A2 complex, ESK1, was used as a negative control for HPV mAb, but positive assay control for RMF/HLA-A2 complex. In parallel, HLA-A2 expression stabilization was determined by staining the cells with anti-HLA-A2 mAb BB7 clone [B]. Binding potency of the mAbs was measured by titrating the HPV-E7p11-19 peptide at the indicated concentrations onto T2 cells; the cells were stained with indicated mAbs at 3ug/ml [C]. Mab titration was performed for relative avidity on T2 cells pulsed with HPV-E7p11-19 peptide at 20ug/ml and stained with the indicated mAbs at concentrations ranging from 3ug/ml to 0.03ug/ml] [D]. All binding was determined by flow cytometry and indicated by median fluorescence intensity [MFI].