Investigation of a monoclonal antibody against enterotoxigenic Escherichia coli, expressed as secretory IgA1 and IgA2 in plants
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 facil...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2021-01-01
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| Series: | Gut Microbes |
| Subjects: | |
| Online Access: | http://dx.doi.org/10.1080/19490976.2020.1859813 |
| _version_ | 1818324025286328320 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Audrey Y-H Teh |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-13T11:22:01Z |
| format | Article |
| id | doaj.art-9adbc1d8ed9d4e8786f8aee96cdadbaa |
| institution | Directory Open Access Journal |
| issn | 1949-0976 1949-0984 |
| language | English |
| last_indexed | 2024-12-13T11:22:01Z |
| publishDate | 2021-01-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Gut Microbes |
| spelling | doaj.art-9adbc1d8ed9d4e8786f8aee96cdadbaa2022-12-21T23:48:25ZengTaylor & Francis GroupGut Microbes1949-09761949-09842021-01-0113110.1080/19490976.2020.18598131859813Investigation of a monoclonal antibody against enterotoxigenic Escherichia coli, expressed as secretory IgA1 and IgA2 in plantsAudrey Y-H Teh0Lisa Cavacini1Yue Hu2Ozan S. Kumru3Jian Xiong4David T. Bolick5Sangeeta B. Joshi6Clemens Grünwald-Gruber7Friedrich Altmann8Mark Klempner9Richard L. Guerrant10David B. Volkin11Yang Wang12Julian K-C. Ma13St. George’s University of London, LondonMassBiologics of the University of Massachusetts Medical SchoolUniversity of KansasUniversity of KansasUniversity of KansasUniversity of Virginia School of MedicineUniversity of KansasUniversität Für Bodenkultur WienUniversität Für Bodenkultur WienMassBiologics of the University of Massachusetts Medical SchoolUniversity of Virginia School of MedicineUniversity of KansasMassBiologics of the University of Massachusetts Medical SchoolSt. George’s University of London, LondonPassive 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.http://dx.doi.org/10.1080/19490976.2020.1859813enterotoxigenic escherichia colimonoclonal antibodysecretory igapassive immunizationimmunotherapynicotiana benthamiana |
| spellingShingle | 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 Investigation of a monoclonal antibody against enterotoxigenic Escherichia coli, expressed as secretory IgA1 and IgA2 in plants Gut Microbes enterotoxigenic escherichia coli monoclonal antibody secretory iga passive immunization immunotherapy nicotiana benthamiana |
| title | Investigation of a monoclonal antibody against enterotoxigenic Escherichia coli, expressed as secretory IgA1 and IgA2 in plants |
| title_full | Investigation of a monoclonal antibody against enterotoxigenic Escherichia coli, expressed as secretory IgA1 and IgA2 in plants |
| title_fullStr | Investigation of a monoclonal antibody against enterotoxigenic Escherichia coli, expressed as secretory IgA1 and IgA2 in plants |
| title_full_unstemmed | Investigation of a monoclonal antibody against enterotoxigenic Escherichia coli, expressed as secretory IgA1 and IgA2 in plants |
| title_short | Investigation of a monoclonal antibody against enterotoxigenic Escherichia coli, expressed as secretory IgA1 and IgA2 in plants |
| title_sort | investigation of a monoclonal antibody against enterotoxigenic escherichia coli expressed as secretory iga1 and iga2 in plants |
| topic | enterotoxigenic escherichia coli monoclonal antibody secretory iga passive immunization immunotherapy nicotiana benthamiana |
| url | http://dx.doi.org/10.1080/19490976.2020.1859813 |
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