Advanced Plant-Based Glycan Engineering

Montero-Morales, Laura; Steinkellner, Herta

doi:10.3389/fbioe.2018.00081

Cited by 111 publications

(104 citation statements)

References 59 publications

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“…The glycosylation state of the antibody is crucial for its stability and function (Mastrangeli et al, 2019). In comparison to mammalian cells which have highly heterogeneous glycoforms that may be detrimental for biopharmaceutical production, advances in plant glycoengineering have allowed the production of monoclonal antibodies (mAbs) with more homogenous human-like glycans (Montero-Morales and Steinkellner, 2018). By removing the endogenous plant-specific β1,2-linked xylose and α1,3-linked fucose, a variety of plant-made antibodies have demonstrated improved immune receptor binding and greater potency compared to commercially available antibodies produced in mammalian cells (Zeitlin et al, 2011;Marusic et al, 2018).…”

Section: Introductionmentioning

confidence: 99%

High Level Production of Monoclonal Antibodies Using an Optimized Plant Expression System

Diamos¹,

Hunter²,

Pardhe³

et al. 2020

Front. Bioeng. Biotechnol.

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Biopharmaceuticals are a large and fast-growing sector of the total pharmaceutical market with antibody-based therapeutics accounting for over 100 billion USD in sales yearly. Mammalian cells are traditionally used for monoclonal antibody production, however plant-based expression systems have significant advantages. In this work, we showcase recent advances made in plant transient expression systems using optimized geminiviral vectors that can efficiently produce heteromultimeric proteins. Two, three, or four fluorescent proteins were coexpressed simultaneously, reaching high yields of 3-5 g/kg leaf fresh weight or ∼50% total soluble protein. As a proof-of-concept for this system, various antibodies were produced using the optimized vectors with special focus given to the creation and production of a chimeric broadly neutralizing anti-flavivirus antibody. The variable regions of this murine antibody, 2A10G6, were codon optimized and fused to a human IgG1. Analysis of the chimeric antibody showed that it was efficiently expressed in plants at 1.5 g of antibody/kilogram of leaf tissue, can be purified to near homogeneity by a simple one-step purification process, retains its ability to recognize the Zika virus envelope protein, and potently neutralizes Zika virus. Two other monoclonal antibodies were produced at similar levels (1.2-1.4 g/kg). This technology will be a versatile tool for the production of a wide spectrum of pharmaceutical multi-protein complexes in a fast, powerful, and cost-effective way.

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Section: Introductionmentioning

confidence: 99%

High Level Production of Monoclonal Antibodies Using an Optimized Plant Expression System

Diamos¹,

Hunter²,

Pardhe³

et al. 2020

Front. Bioeng. Biotechnol.

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“…The capacity of plants to carry out glycosylation is an advantage over prokaryotic expression systems; and even in insect and yeast cells, glycosylation capacity is limited (Marsian & Lomonossoff, ). Glycoengineering in all of the available systems aims to increase the production of human‐like glycosylation profiles in recombinant proteins, and the success of this work may determine the success of individual expression systems in the future (Montero‐Morales & Steinkellner, ; Sethuraman & Stadheim, ).…”

Section: Considerations For Therapeutic Protein Production Systemsmentioning

confidence: 99%

Therapeutic recombinant protein production in plants: Challenges and opportunities

Burnett¹,

Burnett

2019

Plants People Planet

164

105

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Societal Impact Statement Therapeutic protein production in plants is an area of great potential for increasing and improving the production of proteins for the treatment or prevention of disease in humans and other animals. There are a number of key benefits of this technique for scientists and society, as well as regulatory challenges that need to be overcome by policymakers. Increased public understanding of the costs and benefits of therapeutic protein production in plants will be instrumental in increasing the acceptance, and thus the medical and veterinary impact, of this approach. Summary Therapeutic recombinant proteins are a powerful tool for combating many diseases which have previously been hard to treat. The most utilized expression systems are Chinese Hamster Ovary cells and Escherichia coli, but all available expression systems have strengths and weaknesses regarding development time, cost, protein size, yield, growth conditions, posttranslational modifications and regulatory approval. The plant industry is well established and growing and harvesting crops is easy and affordable using current infrastructure. Growth conditions are generally simple: sunlight, water, and the addition of cheap, available fertilizers. There are multiple options for plant expression systems, including species, genetic constructs and protein targeting, each best suited to a particular type of therapeutic protein production. Transient expression systems provide a mechanism to rapidly transfect plants and produce therapeutic protein in a matter of weeks, rather than the months it can take for many competing expression systems, while proteins targeted to cereal seeds can be harvested, stored and potentially purified much more easily than in competing systems. Current challenges for plant expression systems include a lack of regulatory approval, environmental containment concerns and nonhuman glycosylation, which may limit the scope of the type of therapeutic proteins that can be manufactured in plants. The specific strengths of plant expression systems could facilitate the production of certain therapeutic proteins quickly and cheaply in the near future.

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“…The prospects for future exploitation of molecular farmed VNPs are literally limitless: essentially anything that produces VNPs that can be expressed in eukaryotic expression systems, can probably also be expressed in plants; the same is true for proteins normally expressed in bacterial expression systems, given that chloroplasts are effectively intracellular bacterial expression systems within plant cells. The cost savings for plant expression compared to conventional fermentation techniques for bacterial, yeast or mammalian cell expression are well documented (Rademacher et al, 2019;Walwyn, Huddy, & Rybicki, 2015); moreover, improvements in plant expression technology such as engineering of human-type glycosylation (Mamedov & Yusibov, 2013;Montero-Morales & Steinkellner, 2018;Strasser, Altmann, & Steinkellner, 2014), design-ofexperiment approaches to extraction and purification (Buyel, 2015;Buyel, Hubbuch, & Fischer, 2016;Buyel, Twyman, & Fischer, 2015), and plant cell-pack approaches for high-throughput, variable scale evaluation of expression in plant cells (Rademacher et al, 2019), have made the use of plants for VNP production far more accessible to a wider sphere of researchers.…”

Section: Future Prospects For Plant-made Vnpsmentioning

confidence: 99%

Plant molecular farming of virus‐like nanoparticles as vaccines and reagents

Rybicki

2019

WIREs Nanomed Nanobiotechnol

100

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The use of plants for the production of virus‐like nanoparticles (VNPs) dates back to separating natural empty capsids of plant viruses from whole virions nearly 70 years ago, through to the present use of transgenic plants or recombinant Agrobacterium tumefaciens and/or plant virus‐derived vectors for the transient expression of engineered viral or other structural proteins in plants—a production system also known as molecular farming. Plant production of heterologous proteins has major advantages in terms of convenience—whole plants are generally used, and processes do not need to be sterile—and cost, as bulk biomass production is significantly cheaper than by any other method. Plant‐made VNPs in current use for nanotechnology include whole virions and naturally occurring empty capsids of plant viruses, and particles made by reassembly of coat protein (CP) purified from virions or by recombinant expression. Engineered VNP‐forming animal or human virus CPs expressed in plants include L1 protein from human papillomaviruses, human norovirus CP, hepatitis B surface and core antigens, influenza virus HA protein and HIV Gag polyprotein forming large enveloped particles by budding, orbi‐ and rotavirus particles that require assembly of four co‐expressed proteins, and polio‐ and foot and mouth disease viruses which require proteolytic processing of a polyprotein precursor to form 4‐component VNPs. Both plant and animal virus‐derived plant‐made VNPs can be used for surface and internal display of heterologous peptides or even whole proteins. A significant recent development has been the production of pseudovirions in plants, comprising plant or animal virus CPs and RNA or DNA pseudogenomes that can be used to deliver nucleic acid payloads into cultured cells or specific tissues or tumors in whole animals. This article is characterized under: Biology‐Inspired Nanomaterials > Protein and Virus‐Based Structures Therapeutic Approaches and Drug Discovery > Emerging Technologies Diagnostic Tools > in vivo Nanodiagnostics and Imaging

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Advanced Plant-Based Glycan Engineering

Cited by 111 publications

References 59 publications

High Level Production of Monoclonal Antibodies Using an Optimized Plant Expression System

High Level Production of Monoclonal Antibodies Using an Optimized Plant Expression System

Therapeutic recombinant protein production in plants: Challenges and opportunities

Plant molecular farming of virus‐like nanoparticles as vaccines and reagents

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