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Humanization and expression of IgG and IgM antibodies in plants as potential diagnostic reagents for Valley Fever

Monoclonal antibodies (mAbs) are important proteins used in many life science applications, from diagnostics to therapeutics. High demand for mAbs for different applications urges the development of rapid and reliable recombinant production platforms. Plants provide a quick and inexpensive system fo...

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Autores principales: Jugler, Collin, Grill, Francisca J., Eidenberger, Lukas, Karr, Timothy L., Grys, Thomas E., Steinkellner, Herta, Lake, Douglas F., Chen, Qiang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9478164/
https://www.ncbi.nlm.nih.gov/pubmed/36119630
http://dx.doi.org/10.3389/fpls.2022.925008
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author Jugler, Collin
Grill, Francisca J.
Eidenberger, Lukas
Karr, Timothy L.
Grys, Thomas E.
Steinkellner, Herta
Lake, Douglas F.
Chen, Qiang
author_facet Jugler, Collin
Grill, Francisca J.
Eidenberger, Lukas
Karr, Timothy L.
Grys, Thomas E.
Steinkellner, Herta
Lake, Douglas F.
Chen, Qiang
author_sort Jugler, Collin
collection PubMed
description Monoclonal antibodies (mAbs) are important proteins used in many life science applications, from diagnostics to therapeutics. High demand for mAbs for different applications urges the development of rapid and reliable recombinant production platforms. Plants provide a quick and inexpensive system for producing recombinant mAbs. Moreover, when paired with an established platform for mAb discovery, plants can easily be tailored to produce mAbs of different isotypes against the same target. Here, we demonstrate that a hybridoma-generated mouse mAb against chitinase 1 (CTS1), an antigen from Coccidioides spp., can be biologically engineered for use with serologic diagnostic test kits for coccidioidomycosis (Valley Fever) using plant expression. The original mouse IgG was modified and recombinantly produced in glycoengineered Nicotiana benthamiana plants via transient expression as IgG and IgM isotypes with human kappa, gamma, and mu constant regions. The two mAb isotypes produced in plants were shown to maintain target antigen recognition to CTS1 using similar reagents as the Food and Drug Administration (FDA)-approved Valley Fever diagnostic kits. As none of the currently approved kits provide antibody dilution controls, humanization of antibodies that bind to CTS1, a major component of the diagnostic antigen preparation, may provide a solution to the lack of consistently reactive antibody controls for Valley Fever diagnosis. Furthermore, our work provides a foundation for reproducible and consistent production of recombinant mAbs engineered to have a specific isotype for use in diagnostic assays.
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spelling pubmed-94781642022-09-17 Humanization and expression of IgG and IgM antibodies in plants as potential diagnostic reagents for Valley Fever Jugler, Collin Grill, Francisca J. Eidenberger, Lukas Karr, Timothy L. Grys, Thomas E. Steinkellner, Herta Lake, Douglas F. Chen, Qiang Front Plant Sci Plant Science Monoclonal antibodies (mAbs) are important proteins used in many life science applications, from diagnostics to therapeutics. High demand for mAbs for different applications urges the development of rapid and reliable recombinant production platforms. Plants provide a quick and inexpensive system for producing recombinant mAbs. Moreover, when paired with an established platform for mAb discovery, plants can easily be tailored to produce mAbs of different isotypes against the same target. Here, we demonstrate that a hybridoma-generated mouse mAb against chitinase 1 (CTS1), an antigen from Coccidioides spp., can be biologically engineered for use with serologic diagnostic test kits for coccidioidomycosis (Valley Fever) using plant expression. The original mouse IgG was modified and recombinantly produced in glycoengineered Nicotiana benthamiana plants via transient expression as IgG and IgM isotypes with human kappa, gamma, and mu constant regions. The two mAb isotypes produced in plants were shown to maintain target antigen recognition to CTS1 using similar reagents as the Food and Drug Administration (FDA)-approved Valley Fever diagnostic kits. As none of the currently approved kits provide antibody dilution controls, humanization of antibodies that bind to CTS1, a major component of the diagnostic antigen preparation, may provide a solution to the lack of consistently reactive antibody controls for Valley Fever diagnosis. Furthermore, our work provides a foundation for reproducible and consistent production of recombinant mAbs engineered to have a specific isotype for use in diagnostic assays. Frontiers Media S.A. 2022-09-02 /pmc/articles/PMC9478164/ /pubmed/36119630 http://dx.doi.org/10.3389/fpls.2022.925008 Text en Copyright © 2022 Jugler, Grill, Eidenberger, Karr, Grys, Steinkellner, Lake and Chen. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Plant Science
Jugler, Collin
Grill, Francisca J.
Eidenberger, Lukas
Karr, Timothy L.
Grys, Thomas E.
Steinkellner, Herta
Lake, Douglas F.
Chen, Qiang
Humanization and expression of IgG and IgM antibodies in plants as potential diagnostic reagents for Valley Fever
title Humanization and expression of IgG and IgM antibodies in plants as potential diagnostic reagents for Valley Fever
title_full Humanization and expression of IgG and IgM antibodies in plants as potential diagnostic reagents for Valley Fever
title_fullStr Humanization and expression of IgG and IgM antibodies in plants as potential diagnostic reagents for Valley Fever
title_full_unstemmed Humanization and expression of IgG and IgM antibodies in plants as potential diagnostic reagents for Valley Fever
title_short Humanization and expression of IgG and IgM antibodies in plants as potential diagnostic reagents for Valley Fever
title_sort humanization and expression of igg and igm antibodies in plants as potential diagnostic reagents for valley fever
topic Plant Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9478164/
https://www.ncbi.nlm.nih.gov/pubmed/36119630
http://dx.doi.org/10.3389/fpls.2022.925008
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