Expression and immunogenicity assessment of a plant-made immunogen targeting the cytotoxic T-lymphocyte associated antigen-4: a possible approach for cancer immunotherapy

Yiemchavee, Sutita; Wong-Arce, Alejandra; Romero-Maldonado, Andrea; Shanmugaraj, Balamurugan; Monsiváis‐Urenda, Adriana; Phoolcharoen, Waranyoo; Rosales‐Mendoza, Sergio

doi:10.1016/j.jbiotec.2021.01.016

Cited by 10 publications

(7 citation statements)

References 30 publications

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“…The use of plant expression systems for the production of pharmaceutically important proteins ( Bulaon et al, 2020 ; Hanittinan et al, 2020 ), vaccines ( Marsian et al, 2017 ; Rosales-Mendoza et al, 2017 ), diagnostic reagents ( Rattanapisit et al, 2021 ), and antibodies ( Kopertekh et al, 2019 ; Hurtado et al, 2020 ; Rattanapisit et al, 2020 ) were documented. In addition, the prior studies reported that the generation of plant-made protective immunogen and therapeutic antibodies ( Dent et al, 2016 ; Rattanapisit et al, 2019a , b ; Nessa et al, 2020 ; Yiemchavee et al, 2021 ).…”

Section: Discussionmentioning

confidence: 99%

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

et al. 2021

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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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Section: Discussionmentioning

confidence: 99%

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

et al. 2021

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“…According to the presented techno-economic model ( Nandi et al., 2016 ), plant-based expression technology is a promising platform for the production of mAbs. Our team ( Rattanapisit et al., 2019 ; Phakham et al., 2021 ; Yiemchavee et al., 2021 ; Phetphoung et al., 2022 ) have previously expressed recombinant immunotherapeutic antigens and antibodies in plants, providing proof of principle for its viability. To attenuate striking disparities and improve access to treatment in impoverished countries, plant-based approach offers a competitive platform for the development of immunomodulatory agents for cancer immunotherapy.…”

Section: Introductionmentioning

confidence: 99%

Antitumor effect of plant-produced anti-CTLA-4 monoclonal antibody in a murine model of colon cancer

Bulaon,

Khorattanakulchai,

Rattanapisit

et al. 2023

Front. Plant Sci.

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Cytotoxic T lymphocyte-associated protein 4 (CTLA-4) is an immune checkpoint regulator exclusively expressed on T cells that obstructs the cell’s effector functions. Ipilimumab (Yervoy®), a CTLA-4 blocking antibody, emerged as a notable breakthrough in modern cancer treatment, showing upfront clinical benefits in multiple carcinomas. However, the exhilarating cost of checkpoint blockade therapy is discouraging and even utmost prominent in developing countries. Thereby, affordability of cancer care has become a point of emphasis in drug development pipelines. Plant expression system blossomed as a cutting-edge platform for rapid, facile to scale-up, and economical production of recombinant therapeutics. Here, we describe the production of an anti-CTLA-4 2C8 antibody in Nicotiana benthamiana. ELISA and bio-layer interferometry were used to analyze antigen binding and binding kinetics. Anticancer responses in vivo were evaluated using knocked-in mice implanted with syngeneic colon tumor. At 4 days post-infiltration, the antibody was transiently expressed in plants with yields of up to 39.65 ± 8.42 μg/g fresh weight. Plant-produced 2C8 binds to both human and murine CTLA-4, and the plant-produced IgG1 also binds to human FcγRIIIa (V158). In addition, the plant-produced 2C8 monoclonal antibody is as effective as Yervoy® in inhibiting tumor growth in vivo. In conclusion, our study underlines the applicability of plant platform to produce functional therapeutic antibodies with promising potential in cancer immunotherapy.

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“…Generally, adaptive immunity produces tumor antigen-specific T-cells that can destroy cancerous cells. Immune checkpoints (ICs), e.g., CTLA-4, PD-L1, PD-1 and others, are receptor-ligand pairs that are efficient to modulate cancer immunity [3][4][5]. However, cancerous cells also use the immune evasion pathway to escape from the host immune system.…”

Section: Introductionmentioning

confidence: 99%

Expression of plant-produced anti-PD-L1 antibody with anoikis sensitizing activity in human lung cancer cells via., suppression on epithelial-mesenchymal transition

Phetphoung

Malla²,

Rattanapisit³

et al. 2022

PLoS ONE

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Immune checkpoint antibodies in cancer treatment are receptor-ligand pairs that modulate cancer immunity. PD-1/PD-L1 pathway has emerged as one of the major targets in cancer immunotherapy. Atezolizumab, the first anti-PD-L1 antibody approved for the treatment of metastatic urothelial, non-small cell lung, small cell lung and triple-negative breast cancers, is produced in Chinese Hamster Ovary (CHO) cells with several limitations i.e., high-production costs, low-capacity yields, and contamination risks. Due to the rapid scalability and low production costs, the transient expression in Nicotiana benthamiana leaves was investigated by co-infiltration of Agrobacterium tumefaciens GV3101 cultures harboring the nucleic acid sequences encoding for Atezolizumab heavy chain and light chain in this study. The transient expression of Atezolizumab in transformed N. benthamiana accumulated up to 86.76 μg/g fresh leaf weight after 6 days of agroinfiltration (OD 600 nm: 0.4) with 1:1 ratio of heavy chain to light chain. The structural and functional characteristics of plant-produced Atezolizumab was compared with commercially available Tecentriq® from CHO cells with similar binding efficacies to PD-L1 receptor. The direct anti-cancer effect of plant-produced anti-PD-L1 was further performed in human lung metastatic cancer cells H460 cultured under detachment condition, demonstrating the activity of anti-PD-L1-antibody on sensitizing anoikis as well as the suppression on anti-apoptosis proteins (Bcl-2 and Mcl-1) and modulation of epithelial to mesenchymal regulating proteins (E-cadherin, N-cadherin, Snail and Slug). In conclusion, this study manifests plants as an alternative cost-effective platform for the production of functional monoclonal antibodies for use in cancer therapy.

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Expression and immunogenicity assessment of a plant-made immunogen targeting the cytotoxic T-lymphocyte associated antigen-4: a possible approach for cancer immunotherapy

Cited by 10 publications

References 30 publications

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

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

Antitumor effect of plant-produced anti-CTLA-4 monoclonal antibody in a murine model of colon cancer

Expression of plant-produced anti-PD-L1 antibody with anoikis sensitizing activity in human lung cancer cells via., suppression on epithelial-mesenchymal transition

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