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. 2014 Jun 24;289(32):22035–22047. doi: 10.1074/jbc.M114.568725

FIGURE 5.

FIGURE 5.

scFv clone D12 could recognize PBF A2.2 peptide presented on the surface of sarcoma cells. A and B, FACS analysis of reactivity of D12 scFv-hIgG1 (1 mg/ml) and D12 scFv multimer (10 μg/ml) against T2 cells pulsed with the indicated peptides. %MFI increase is indicated. T2 without peptide was used as the negative control. A more than 20.0% MFI increase is indicated by underlining. C, FACS analysis of the reactivity of the D12 scFv multimer (3 μg/ml) with sarcoma cell lines and primary culture cells. Osteosarcoma cell line U2OS (A*02:01/A*3201, PBF+), OS2000 (A*2402, PBF+), KIKU (A*0206/A*2402, PBF+), HOS (A*02:11, PBF+), Saos-2 (A*02:01/A*24:02, PBF+), malignant fibrous histiocytoma cell line MFH2003 (A*2402, PBF−), and primary culture of osteosarcoma (Primary OS; A*02:01, PBF+) were used as target cells. A more than 20.0% MFI increase is indicated by underlining. The expression status of HLA-A2·PBF A2.2 peptide complex was graded as follows: strong (≥100% MFI increase), weak (≥20% but <100% MFI increase), and none (<20% MFI increase).