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The Binding Ability and Binding Stability between OVA66 Peptides and HLA-A*0201 Molecule
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Abstract:
Objective: To establish the methods for screening the HLA-A2-restricted peptides on T2-dependent cell model. Methods: The binding ability and binding stability of peptides to HLA-A2 molecule was determined by measuring peptide-induced expression of HLA-A2 molecules on TAP-deficient cell line-T2 cells. Briefly, T2 cells were incubated with OVA66 candidate peptides at different concentration(0.2 μg/ml, 2 μg/ml, 5 μg/ml, 20 μg/ml) and different time. After incubation, expression of HLA-A2 molecules on T2 cells was detected by flow cytometry with murine mAb BB7.2 against human HLA-A2 molecule. The binding ability of peptides was calculated with peptide concentration of 20% MFImax, while the binding stability of peptides was calculated with percentage of MFI(4h)/MFI(0 h).Results: Four OVA66 candidate peptides were assayed with this method. Compared with reference peptide, two peptides (L235 and L238) displayed strong binding ability and good stability, while other two peptides (L236 and L237) exhibited low affinity to HLA-A2 molecules on T2 cells. Conclusion: With T2-dependent cell model, we could test the binding ability and binding stability of peptides before inducing peptide-specific T cell lines in vitro.
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