Rapid Transient Production of a Monoclonal Antibody Neutralizing the Porcine Epidemic Diarrhea Virus (PEDV) in Nicotiana benthamiana and Lactuca sativa

Rattanapisit, Kaewta; Srijangwad, Anchalee; Chuanasa, Taksina; Sukrong, Suchada; Tantituvanont, Angkana; Mason, Hugh S.; Nilubol, Dachrit; Phoolcharoen, Waranyoo

doi:10.1055/s-0043-112344

Cited by 8 publications

(12 citation statements)

References 43 publications

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“…Previous studies have described tobacco-derived plantibodies that showed potent therapeutic activity against multiple viruses such as EBOV, HBV, HIV, PEDV, RABV, WNV, and others in small animal models of infection, e.g., mice [39,[42][43][44][45][46]. However, to our knowledge, this is the first study demonstrating the ability of plantibodies to protect against influenza viral infections.…”

Section: Discussionmentioning

confidence: 86%

“…Moreover, plantibodies overcome some of the concerns associated with animal-derived therapeutic MAbs obtained from serum or plasma, including intermediate reactions, pyrogenicity, potential contamination with other zoonotic pathogens and/or toxins, and serum sickness [32,[34][35][36][37]. For these reasons, plantibodies have been proposed for the treatment of several bacterial (e.g., Salmonella, Streptococcus, Porphyromonas) and viral (e.g., Ebola virus, EBOV; Hepatitis B virus, HBV; Human immunodeficiency virus, HIV; Porcine epidemic diarrhea virus, PEDV; Rabies virus, RABV; West Nile virus, WNV) infections or toxins (e.g., ricin and shiga toxin) [35,[38][39][40][41][42][43][44][45][46]. However, to date, plantibodies have not been used for the treatment of influenza viral infections.…”

Section: Introductionmentioning

confidence: 99%

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A Broad and Potent H1-Specific Human Monoclonal Antibody Produced in Plants Prevents Influenza Virus Infection and Transmission in Guinea Pigs

Park

Piepenbrink

et al. 2020

Viruses

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Although seasonal influenza vaccines block most predominant influenza types and subtypes, humans still remain vulnerable to waves of seasonal and new potential pandemic influenza viruses for which no immunity may exist because of viral antigenic drift and/or shift. Previously, we described a human monoclonal antibody (hMAb), KPF1, which was produced in human embryonic kidney 293T cells (KPF1-HEK) with broad and potent neutralizing activity against H1N1 influenza A viruses (IAV) in vitro, and prophylactic and therapeutic activities in vivo. In this study, we produced hMAb KPF1 in tobacco plants (KPF1-Antx) and demonstrated how the plant-produced KPF1-Antx hMAb possesses similar biological activity compared with the mammalian-produced KPF1-HEK hMAb. KPF1-Antx hMAb showed broad binding to recombinant HA proteins and H1N1 IAV, including A/California/04/2009 (pH1N1) in vitro, which was comparable to that observed with KPF1-HEK hMAb. Importantly, prophylactic administration of KPF1-Antx hMAb to guinea pigs prevented pH1N1 infection and transmission in both prophylactic and therapeutic experiments, substantiating its clinical potential to prevent and treat H1N1 infections. Collectively, this study demonstrated, for the first time, a plant-produced influenza hMAb with in vitro and in vivo activity against influenza virus. Because of the many advantages of plant-produced hMAbs, such as rapid batch production, low cost, and the absence of mammalian cell products, they represent an alternative strategy for the production of immunotherapeutics for the treatment of influenza viral infections, including emerging seasonal and/or pandemic strains.

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Section: Discussionmentioning

confidence: 86%

Section: Introductionmentioning

confidence: 99%

A Broad and Potent H1-Specific Human Monoclonal Antibody Produced in Plants Prevents Influenza Virus Infection and Transmission in Guinea Pigs

Park

Piepenbrink

et al. 2020

Viruses

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“…Efficient complex formation is required to produce antibodies in plant cells. It has previously been shown that non-competing amplification of two different replicons can be achieved using a geminiviral replication vector (Phoolcharoen et al, 2011;Diamos et al, 2016;Rattanapisit et al, 2017). The most renowned and widely used transient expression system is the magnICON system (Marillonnet et al, 2005).…”

Section: Discussionmentioning

confidence: 99%

RAP Tag and PMab-2 Antibody: A Tagging System for Detecting and Purifying Proteins in Plant Cells

et al. 2020

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An affinity tag system requires both high affinity and specificity. The RAP tag epitope DMVNPGLEDRIE, derived from rat podoplanin (PDPN), is specifically recognized by PMab-2 monoclonal antibodies in rats. Here, we demonstrated that high levels of PMab-2 can be produced in Nicotiana benthamiana and plant-derived PMab-2 possesses similar activity to CHO-derived PMab-2, and the RAP tag presents a useful tagging system for detecting and purifying proteins from plant cells. The heavy chain of PMab-2 fused with KDEL, an endoplasmic reticulum retention sequence, and the light chain of the antibody were introduced into N. benthamiana by agroinfiltration. The expression of PMab-2 peaked 4 days after agroinfiltration, and approximately 0.3 mg/g fresh weight of the antibody was accumulated. After purification, the plant-derived PMab-2 successfully recognized rat PDPN expressed in CHO-K1 cells and exhibited almost the same binding activity as CHO-derived PMab-2. The RAP-tagged proteins expressed in plant cells were specifically recognized by PMab-2. These results indicate that PMab-2 can accumulate at high levels in N. benthamiana and is easily purified and that the RAP tagging system presents a useful tool for detecting and purifying proteins of interest in plant cells.

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“…Plants have been used to produce a variety of recombinant proteins, such as monoclonal antibodies, vaccines, therapeutic proteins, enzymes, etc. [18][19][20][21].…”

Section: Discussionmentioning

confidence: 99%

Recombinant Human Dentin Matrix Protein 1 (hDMP1) Expressed in Nicotiana benthamiana Potentially Induces Osteogenic Differentiation

Ahmad

Kaewpungsup

Khorattanakulchai

et al. 2019

Plants

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Inductive molecules are critical components for successful bone tissue engineering. Dentin matrix protein-1 (DMP1), a non-collagenous protein in the bone matrix, has been shown to play roles in osteogenic differentiation and phosphate homeostasis. This study aimed to produce recombinant human dentin matrix protein-1 (hDMP1) in Nicotiana benthamiana and investigated the ability of this plant-produced DMP1 to induce osteogenesis in human periodontal ligament stem cells (hPDLSCs). The hDMP1 gene was cloned into the geminiviral vector for transient expression in N. benthamiana. We found that hDMP1 was transiently expressed in N. benthamiana leaves and could be purified by ammonium sulphate precipitation followed by nickel affinity chromatography. The effects of hDMP1 on the induction of cell proliferation and osteogenic differentiation were investigated. The results indicated that plant-produced hDMP1 could induce the cell proliferation of hPDLSCs and increase the expression levels of osteogenic genes, including osterix (OSX), type I collagen (COL1), bone morphogenetic protein-2 (BMP2), and Wnt3a. Moreover, the plant-produced hDMP1 promoted calcium deposition in hPDLSCs as determined by alizarin red S staining. In conclusion, our results indicated that plant-produced hDMP1 could induce osteogenic differentiation in hPDLSCs and could potentially be used as a bone inducer in bone tissue engineering.

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Rapid Transient Production of a Monoclonal Antibody Neutralizing the Porcine Epidemic Diarrhea Virus (PEDV) in Nicotiana benthamiana and Lactuca sativa

Cited by 8 publications

References 43 publications

A Broad and Potent H1-Specific Human Monoclonal Antibody Produced in Plants Prevents Influenza Virus Infection and Transmission in Guinea Pigs

A Broad and Potent H1-Specific Human Monoclonal Antibody Produced in Plants Prevents Influenza Virus Infection and Transmission in Guinea Pigs

RAP Tag and PMab-2 Antibody: A Tagging System for Detecting and Purifying Proteins in Plant Cells

Recombinant Human Dentin Matrix Protein 1 (hDMP1) Expressed in Nicotiana benthamiana Potentially Induces Osteogenic Differentiation

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