A Dual-Approach Strategy to Optimize the Safety and Efficacy of Anti-Zika Virus Monoclonal Antibody Therapeutics

Abstract

Antibody-dependent enhancement of infection (ADE) is clinically relevant to Dengue virus (DENV) infection and poses a major risk to the application of monoclonal antibody (mAb)-based therapeutics against related flaviviruses such as the Zika virus (ZIKV). Here, we tested a two-tier approach for selecting non-cross-reactive mAbs combined with modulating Fc glycosylation as a strategy to doubly secure the elimination of ADE while preserving Fc effector functions. To this end, we selected a ZIKV-specific mAb (ZV54) and generated three ZV54 variants using Chinese hamster ovary cells and wild-type (WT) and glycoengineered ΔXF Nicotiana benthamiana plants as production hosts (ZV54CHO, ZV54WT, and ZV54ΔXF). The three ZV54 variants shared an identical polypeptide backbone, but each exhibited a distinct Fc N-glycosylation profile. All three ZV54 variants showed similar neutralization potency against ZIKV but no ADE activity for DENV infection, validating the importance of selecting the virus/serotype-specific mAbs for avoiding ADE by related flaviviruses. For ZIKV infection, however, ZV54CHO and ZV54ΔXF showed significant ADE activity while ZV54WT completely forwent ADE, suggesting that Fc glycan modulation may yield mAb glycoforms that abrogate ADE even for homologous viruses. In contrast to the current strategies for Fc mutations that abrogate all effector functions along with ADE, our approach allowed the preservation of effector functions as all ZV54 glycovariants retained antibody-dependent cellular cytotoxicity (ADCC) against the ZIKV-infected cells. Furthermore, the ADE-free ZV54WT demonstrated in vivo efficacy in a ZIKV-infection mouse model. Collectively, our study provides further support for the hypothesis that antibody–viral surface antigen and Fc-mediated host cell interactions are both prerequisites for ADE, and that a dual-approach strategy, as shown herein, contributes to the development of highly safe and efficacious anti-ZIKV mAb therapeutics. Our findings may be impactful to other ADE-prone viruses, including SARS-CoV-2.

A Review: Understanding Molecular Mechanisms of Antibody-Dependent Enhancement in Viral Infections

“…In the treatment of monoclonal antibodies, the modulation of Fc glycosylation has been demonstrated to be a useful method for preventing ADE since it preserves the effector function, which is lost in Fc mutation methods. These discoveries may have an influence on ADE-prone viruses like SARS-CoV-2 [ 76 ].…”

A Monoclonal Antibody Produced in Glycoengineered Plants Potently Neutralizes Monkeypox Virus

“…Our results show the rapid expression of 7D11 in N. benthamiana plants with a peak accumulation level at 642 µg/gram LFW 6-9 days posts DNA construct delivery. Compared with other mAbs that were recently developed by our group using the same plant-expression vector [19,32], the accumulation level of p7D11 mAb in leaves is much higher, indicating its potential for potential large-scale production. Published reports on the expression levels of 7D11 mAb in non-plant-based systems are currently not available.…”

“…Our results show the rapid expression of 7D11 plants with a peak accumulation level at 642 µg/gram LFW 6-9 days pos delivery. Compared with other mAbs that were recently developed by the same plant-expression vector [19,32], the accumulation level of p7D Figure 6. Neutralization of Monkeypox virus by p7D11 mAb.…”

“…Indirect ELISA was used to investigate the specific binding of p7D11 mAb to the MPXV L1 protein (MedChemExpress, Monmouth Junction, NJ, USA) using a previously described method [32]. Briefly, 2 µg/mL of L1 was coated on 96-well plates overnight at 4 • C and blocked with 5% milk in 1x PBST.…”

“…We characterized the N-glycosylation population of 7D11 mAb using mass spectrometry with our previously published procedures [23,32]. Briefly, 7D11 mAb was separated on SDS-PAGE, and HC was excised and digested with trypsin to generate glycopeptides, which were further identified by a liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS) system (Orbitrap Exploris 480, Thermo Scientific, Waltham, MA, USA).…”