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Investigation of a monoclonal antibody against enterotoxigenic Escherichia coli, expressed as secretory IgA1 and IgA2 in plants
Teh, Audrey Y-H Cavacini, Lisa Hu, Yue Kumru, Ozan S. Xiong, Jian Bolick, David T. Joshi, Sangeeta B. Grünwald-Gruber, Clemens Altmann, Friedrich Klempner, Mark
Teh, Audrey Y-H
Cavacini, Lisa
Hu, Yue
Kumru, Ozan S.
Xiong, Jian
Bolick, David T.
Joshi, Sangeeta B.
Grünwald-Gruber, Clemens
Altmann, Friedrich
Klempner, Mark
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Abstract
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.
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Date
2021-01-13
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Taylor & Francis Open Access
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Keywords
Enterotoxigenic Escherichia coli, Monoclonal antibody, Secretory IgA, Passive immunization, Immunotherapy, Nicotiana benthamiana
Citation
Audrey Y-H Teh, Lisa Cavacini, Yue Hu, Ozan S. Kumru, Jian Xiong, David T. Bolick, Sangeeta B. Joshi, Clemens Grünwald-Gruber, Friedrich Altmann, Mark Klempner, Richard L. Guerrant, David B. Volkin, Yang Wang & Julian K-C. Ma (2021) Investigation of a monoclonal antibody against enterotoxigenic Escherichia coli, expressed as secretory IgA1 and IgA2 in plants, Gut Microbes, 13:1, 1859813, DOI: 10.1080/19490976.2020.1859813