Amino Acids, Peptides, and Proteins | Bacteria | Bacterial Infections and Mycoses | Biological Factors | Immunotherapy
Passive immunization with antibodies is a promising approach against enterotoxigenic Escherichia coli diarrhea, a prevalent disease in LMICs. The objective of this study was to investigate expression of a monoclonal anti-ETEC CfaE secretory IgA antibody in N. benthamiana plants, with a view to facilitating access to ETEC passive immunotherapy. SIgA1 and SIgA2 forms of mAb 68-81 were produced by co-expressing the light and engineered heavy chains with J chain and secretory component in N. benthamiana. Antibody expression and assembly were compared with CHO-derived antibodies by SDS-PAGE, western blotting, size-exclusion chromatography and LC-MS peptide mapping. N-linked glycosylation was assessed by rapid fluorescence/mass spectrometry and LC-ESI-MS. Susceptibility to gastric digestion was assessed in an in vitro model. Antibody function was compared for antigen binding, a Caco-2 cell-based ETEC adhesion assay, an ETEC hemagglutination inhibition assay and a murine in vivo challenge study. SIgA1 assembly appeared superior to SIgA2 in plants. Both sub-classes exhibited resistance to degradation by simulated gastric fluid, comparable to CHO-produced 68-61 SIgA1. The plant expressed SIgAs had more homogeneous N-glycosylation than CHO-derived SIgAs, but no alteration of in vitro functional activity was observed, including antibodies expressed in a plant line engineered for mammalian-like N glycosylation. The plant-derived SIgA2 mAb demonstrated protection against diarrhea in a murine infection model. Although antibody yield and purification need to be optimized, anti-ETEC SIgA antibodies produced in a low-cost plant platform are functionally equivalent to CHO antibodies, and provide promise for passive immunotherapy in LMICs.
Enterotoxigenic Escherichia coli, Nicotiana benthamiana, immunotherapy, monoclonal antibody, passive immunization, secretory IgA
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DOI of Published Version
Teh AY, Cavacini L, Hu Y, Kumru OS, Xiong J, Bolick DT, Joshi SB, Grünwald-Gruber C, Altmann F, Klempner M, Guerrant RL, Volkin DB, Wang Y, Ma JK. Investigation of a monoclonal antibody against enterotoxigenic Escherichia coli, expressed as secretory IgA1 and IgA2 in plants. Gut Microbes. 2021 Jan-Dec;13(1):1-14. doi: 10.1080/19490976.2020.1859813. PMID: 33439092; PMCID: PMC7833773. Link to article on publisher's site
Teh AY, Cavacini LA, Hu Y, Kumru OS, Xiong J, Bolick DT, Joshi SB, Grunwald-Gruber C, Altmann F, Klempner MS, Guerrant RL, Volkin DB, Wang Y, Ma JK. (2021). Investigation of a monoclonal antibody against enterotoxigenic Escherichia coli, expressed as secretory IgA1 and IgA2 in plants. Open Access Publications by UMMS Authors. https://doi.org/10.1080/19490976.2020.1859813. Retrieved from https://escholarship.umassmed.edu/oapubs/4540
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