Identification of a monoclonal antibody that targets PD-1 in a manner requiring PD-1 Asn58 glycosylation

Mingzhu Wang,Jinchao Zhang,Min Zhang,Shasha Jiao,Shuang Wang,Rongjuan Wang,Junchao Wang

doi:10.1038/s42003-019-0642-9

Mingzhu Wang, Jinchao Zhang + Show 5 more

Open Access

https://doi.org/10.1038/s42003-019-0642-9

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Abstract

Programmed cell death 1 (PD-1) is inhibitory receptor and immune checkpoint protein. Blocking the interaction of PD-1 and its ligands PD-L1/ L2 is able to active T-cell-mediated antitumor response. Monoclonal antibody-based drugs targeting PD-1 pathway have exhibited great promise in cancer therapy. Here we show that MW11-h317, an anti-PD-1 monoclonal antibody, displays high affinity for PD-1 and blocks PD-1 interactions with PD-L1/L2. MW11-h317 can effectively induce T-cell-mediated immune response and inhibit tumor growth in mouse model. Crystal structure of PD-1/MW11-h317 Fab complex reveals that both the loops and glycosylation of PD-1 are involved in recognition and binding, in which Asn58 glycosylation plays a critical role. The unique glycan epitope in PD-1 to MW11-h317 is different from the first two approved clinical PD-1 antibodies, nivolumab and pembrolizumab. These results suggest MW11-h317 as a therapeutic monoclonal antibody of PD-1 glycosylation-targeting which may become efficient alternative for cancer therapy.

Highlights

Programmed cell death 1 (PD-1) is inhibitory receptor and immune checkpoint protein
We immunized mice with the recombinant human PD-1 protein and identified the murine monoclonal antibody m317 that bound to human PD-1 protein and inhibited interactions of PD-1 and ligands from hybridoma
The results revealed that N58A-mutated PD-1 exhibited significant loss of MW11-h317 binding, while the other three mutants (N49A, N74, and N116) displayed no changes compared with WT protein (Fig. 4)

Summary

Introduction

Programmed cell death 1 (PD-1) is inhibitory receptor and immune checkpoint protein. Blocking the interaction of PD-1 and its ligands PD-L1/ L2 is able to active T-cell-mediated antitumor response. Nivolumab and pembrolizumab, belong to the IgG4 subclass, and their binding depends on the interactions with the flexible loops of PD-1, and the epitopes include residues in the PD-L1-binding site. Structural analysis revealed that the loops of PD-1 contributed to the interaction with MW11-h317 Fab (Fig. 2a).

Results

Conclusion