【摘 要】
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PD-1 plays an important role as an immune checkpoint.Sintilimab is a newly approved PD-1 antibody for cancer immunotherapy with an unknown binding epitope on PD
【机 构】
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School of Biomedical Engineering,Shanghai Jiao Tong University,Shanghai 200240,China;Key Laboratory
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PD-1 plays an important role as an immune checkpoint.Sintilimab is a newly approved PD-1 antibody for cancer immunotherapy with an unknown binding epitope on PD-1.In this study,to elucidate the molecular mechanism by which sintilimab blocks PD-1 activation,we applied Anti-body binding epitope Mapping(AbMap)to identify the binding epitope of sintilimab.An epitope was successfully identified,i.e.SLAPKA,aa 127-132.By constructing a series of point mutations,the dominant residues S127,L128,A129,P130,and A132 of PD-1 were further validated by western blot analysis,biolayer interferometry,and flow cytometry.Structural analysis showed that the epitope is partially within the binding interface of PD-1 and PD-L1,and this epitope also partially overlaps with that of nivolumab and pembrolizumab.These results demonstrate that sintilimab can attenuate PD-1 activation by directly competing with the interaction between PD-1 and PD-L1 through binding with the key residues of the FG loop on PD-1.This study also demonstrates the high efficiency and accuracy of AbMap for determining the binding epitope of therapeutic antibodies.
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