Teh, AY-H;
Cavacini, L;
Hu, Y;
Kumru, OS;
Xiong, J;
Bolick, DT;
Joshi, SB;
Grünwald-Gruber, C;
Altmann, F;
Klempner, M;
et al.
Teh, AY-H; 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-C
(2021)
Investigation of a monoclonal antibody against enterotoxigenic Escherichia coli, expressed as secretory IgA1 and IgA2 in plants.
Gut Microbes, 13 (1).
pp. 1-14.
ISSN 1949-0984
https://doi.org/10.1080/19490976.2020.1859813
SGUL Authors: Ma, Julian Teh, Yi Hui
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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.
Item Type: | Article | ||||||||||||||||||
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Additional Information: | © 2021 The Author(s). Published with license by Taylor & Francis Group, LLC. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. | ||||||||||||||||||
Keywords: | Enterotoxigenic Escherichia coli, Nicotiana benthamiana, immunotherapy, monoclonal antibody, passive immunization, secretory IgA | ||||||||||||||||||
SGUL Research Institute / Research Centre: | Academic Structure > Infection and Immunity Research Institute (INII) | ||||||||||||||||||
Journal or Publication Title: | Gut Microbes | ||||||||||||||||||
ISSN: | 1949-0984 | ||||||||||||||||||
Language: | eng | ||||||||||||||||||
Publisher License: | Creative Commons: Attribution 4.0 | ||||||||||||||||||
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PubMed ID: | 33439092 | ||||||||||||||||||
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URI: | https://openaccess.sgul.ac.uk/id/eprint/112681 | ||||||||||||||||||
Publisher's version: | https://doi.org/10.1080/19490976.2020.1859813 |
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