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dc.contributor.authorTeh, Audrey Y-H.
dc.contributor.authorCavacini, Lisa A.
dc.contributor.authorHu, Yue
dc.contributor.authorKumru, Ozan S.
dc.contributor.authorXiong, Jian
dc.contributor.authorBolick, David T.
dc.contributor.authorJoshi, Sangeeta B.
dc.contributor.authorGrunwald-Gruber, Clemens
dc.contributor.authorAltmann, Friedrich
dc.contributor.authorKlempner, Mark S.
dc.contributor.authorGuerrant, Richard L.
dc.contributor.authorVolkin, David B.
dc.contributor.authorWang, Yang
dc.contributor.authorMa, Julian K-C.
dc.date2022-08-11T08:09:58.000
dc.date.accessioned2022-08-23T16:50:53Z
dc.date.available2022-08-23T16:50:53Z
dc.date.issued2021-01-13
dc.date.submitted2021-03-11
dc.identifier.citation<p>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 <em>Escherichia coli</em>, 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. <a href="https://doi.org/10.1080/19490976.2020.1859813">Link to article on publisher's site</a></p>
dc.identifier.issn1949-0976 (Linking)
dc.identifier.doi10.1080/19490976.2020.1859813
dc.identifier.pmid33439092
dc.identifier.urihttp://hdl.handle.net/20.500.14038/41746
dc.description.abstractPassive 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.
dc.language.isoen_US
dc.relation<p><a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&list_uids=33439092&dopt=Abstract">Link to Article in PubMed</a></p>
dc.rights© 2021 The Author(s). 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.
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subjectEnterotoxigenic Escherichia coli
dc.subjectNicotiana benthamiana
dc.subjectimmunotherapy
dc.subjectmonoclonal antibody
dc.subjectpassive immunization
dc.subjectsecretory IgA
dc.subjectAmino Acids, Peptides, and Proteins
dc.subjectBacteria
dc.subjectBacterial Infections and Mycoses
dc.subjectBiological Factors
dc.subjectImmunotherapy
dc.titleInvestigation of a monoclonal antibody against enterotoxigenic Escherichia coli, expressed as secretory IgA1 and IgA2 in plants
dc.typeJournal Article
dc.source.journaltitleGut microbes
dc.source.volume13
dc.source.issue1
dc.identifier.legacyfulltexthttps://escholarship.umassmed.edu/cgi/viewcontent.cgi?article=5571&amp;context=oapubs&amp;unstamped=1
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/oapubs/4540
dc.identifier.contextkey22022754
refterms.dateFOA2022-08-23T16:50:53Z
html.description.abstract<p>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.</p>
dc.identifier.submissionpathoapubs/4540
dc.contributor.departmentMassBiologics
dc.source.pages1-14


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© 2021 The Author(s). 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.
Except where otherwise noted, this item's license is described as © 2021 The Author(s). 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.