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Functional Characterization of Pembrolizumab Produced in Nicotiana benthamiana Using a Rapid Transient Expression System

The striking innovation and clinical success of immune checkpoint inhibitors (ICIs) have undoubtedly contributed to a breakthrough in cancer immunotherapy. Generally, ICIs produced in mammalian cells requires high investment, production costs, and involves time consuming procedures. Recently, the pl...

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Autores principales: Phakham, Tanapati, Bulaon, Christine Joy I., Khorattanakulchai, Narach, Shanmugaraj, Balamurugan, Buranapraditkun, Supranee, Boonkrai, Chatikorn, Sooksai, Sarintip, Hirankarn, Nattiya, Abe, Yoshito, Strasser, Richard, Rattanapisit, Kaewta, Phoolcharoen, Waranyoo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8459022/
https://www.ncbi.nlm.nih.gov/pubmed/34567049
http://dx.doi.org/10.3389/fpls.2021.736299
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author Phakham, Tanapati
Bulaon, Christine Joy I.
Khorattanakulchai, Narach
Shanmugaraj, Balamurugan
Buranapraditkun, Supranee
Boonkrai, Chatikorn
Sooksai, Sarintip
Hirankarn, Nattiya
Abe, Yoshito
Strasser, Richard
Rattanapisit, Kaewta
Phoolcharoen, Waranyoo
author_facet Phakham, Tanapati
Bulaon, Christine Joy I.
Khorattanakulchai, Narach
Shanmugaraj, Balamurugan
Buranapraditkun, Supranee
Boonkrai, Chatikorn
Sooksai, Sarintip
Hirankarn, Nattiya
Abe, Yoshito
Strasser, Richard
Rattanapisit, Kaewta
Phoolcharoen, Waranyoo
author_sort Phakham, Tanapati
collection PubMed
description The striking innovation and clinical success of immune checkpoint inhibitors (ICIs) have undoubtedly contributed to a breakthrough in cancer immunotherapy. Generally, ICIs produced in mammalian cells requires high investment, production costs, and involves time consuming procedures. Recently, the plants are considered as an emerging protein production platform due to its cost-effectiveness and rapidity for the production of recombinant biopharmaceuticals. This study explored the potential of plant-based system to produce an anti-human PD-1 monoclonal antibody (mAb), Pembrolizumab, in Nicotiana benthamiana. The transient expression of this mAb in wild-type N. benthamiana accumulated up to 344.12 ± 98.23 μg/g fresh leaf weight after 4 days of agroinfiltration. The physicochemical and functional characteristics of plant-produced Pembrolizumab were compared to mammalian cell-produced commercial Pembrolizumab (Keytruda(®)). Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and western blot analysis results demonstrated that the plant-produced Pembrolizumab has the expected molecular weight and is comparable with the Keytruda(®). Structural characterization also confirmed that both antibodies have no protein aggregation and similar secondary and tertiary structures. Furthermore, the plant-produced Pembrolizumab displayed no differences in its binding efficacy to PD-1 protein and inhibitory activity between programmed cell death 1 (PD-1) and programmed cell death ligand 1 (PD-L1) interaction with the Keytruda(®). In vitro efficacy for T cell activation demonstrated that the plant-produced Pembrolizumab could induce IL-2 and IFN-γ production. Hence, this proof-of-concept study showed that the plant-production platform can be utilized for the rapid production of functional mAbs for immunotherapy.
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spelling pubmed-84590222021-09-24 Functional Characterization of Pembrolizumab Produced in Nicotiana benthamiana Using a Rapid Transient Expression System Phakham, Tanapati Bulaon, Christine Joy I. Khorattanakulchai, Narach Shanmugaraj, Balamurugan Buranapraditkun, Supranee Boonkrai, Chatikorn Sooksai, Sarintip Hirankarn, Nattiya Abe, Yoshito Strasser, Richard Rattanapisit, Kaewta Phoolcharoen, Waranyoo Front Plant Sci Plant Science The striking innovation and clinical success of immune checkpoint inhibitors (ICIs) have undoubtedly contributed to a breakthrough in cancer immunotherapy. Generally, ICIs produced in mammalian cells requires high investment, production costs, and involves time consuming procedures. Recently, the plants are considered as an emerging protein production platform due to its cost-effectiveness and rapidity for the production of recombinant biopharmaceuticals. This study explored the potential of plant-based system to produce an anti-human PD-1 monoclonal antibody (mAb), Pembrolizumab, in Nicotiana benthamiana. The transient expression of this mAb in wild-type N. benthamiana accumulated up to 344.12 ± 98.23 μg/g fresh leaf weight after 4 days of agroinfiltration. The physicochemical and functional characteristics of plant-produced Pembrolizumab were compared to mammalian cell-produced commercial Pembrolizumab (Keytruda(®)). Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and western blot analysis results demonstrated that the plant-produced Pembrolizumab has the expected molecular weight and is comparable with the Keytruda(®). Structural characterization also confirmed that both antibodies have no protein aggregation and similar secondary and tertiary structures. Furthermore, the plant-produced Pembrolizumab displayed no differences in its binding efficacy to PD-1 protein and inhibitory activity between programmed cell death 1 (PD-1) and programmed cell death ligand 1 (PD-L1) interaction with the Keytruda(®). In vitro efficacy for T cell activation demonstrated that the plant-produced Pembrolizumab could induce IL-2 and IFN-γ production. Hence, this proof-of-concept study showed that the plant-production platform can be utilized for the rapid production of functional mAbs for immunotherapy. Frontiers Media S.A. 2021-09-09 /pmc/articles/PMC8459022/ /pubmed/34567049 http://dx.doi.org/10.3389/fpls.2021.736299 Text en Copyright © 2021 Phakham, Bulaon, Khorattanakulchai, Shanmugaraj, Buranapraditkun, Boonkrai, Sooksai, Hirankarn, Abe, Strasser, Rattanapisit and Phoolcharoen. 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
Phakham, Tanapati
Bulaon, Christine Joy I.
Khorattanakulchai, Narach
Shanmugaraj, Balamurugan
Buranapraditkun, Supranee
Boonkrai, Chatikorn
Sooksai, Sarintip
Hirankarn, Nattiya
Abe, Yoshito
Strasser, Richard
Rattanapisit, Kaewta
Phoolcharoen, Waranyoo
Functional Characterization of Pembrolizumab Produced in Nicotiana benthamiana Using a Rapid Transient Expression System
title Functional Characterization of Pembrolizumab Produced in Nicotiana benthamiana Using a Rapid Transient Expression System
title_full Functional Characterization of Pembrolizumab Produced in Nicotiana benthamiana Using a Rapid Transient Expression System
title_fullStr Functional Characterization of Pembrolizumab Produced in Nicotiana benthamiana Using a Rapid Transient Expression System
title_full_unstemmed Functional Characterization of Pembrolizumab Produced in Nicotiana benthamiana Using a Rapid Transient Expression System
title_short Functional Characterization of Pembrolizumab Produced in Nicotiana benthamiana Using a Rapid Transient Expression System
title_sort functional characterization of pembrolizumab produced in nicotiana benthamiana using a rapid transient expression system
topic Plant Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8459022/
https://www.ncbi.nlm.nih.gov/pubmed/34567049
http://dx.doi.org/10.3389/fpls.2021.736299
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