ArticlePloS one2013
Protection of recombinant mammalian antibodies from development-dependent proteolysis in leaves of Nicotiana benthamiana.
Article in PloS one, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 29 papers.
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Who cites it
29 citing papers in PubMed.
- Gene Editing of Nicotiana benthamiana Architecture for Space-Efficient Production of Recombinant Proteins in Closed Environments.Plant biotechnology journal · 2026Article
- Nicotiana benthamiana's Responses to Agroinfiltration, a Treasure Grove of New Avenues to Improve Protein Yields in Plant Molecular Farming.Plant biotechnology journal · 2026Review
- Plant-Based Ligand-Depletion Binding Assay for Assessing Pattern Recognition Receptor-Ligand Interactions.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Rhizobium rhizogenes A4-derived strains mediate hyper-efficient transient gene expression in Nicotiana benthamiana and other solanaceous plants.Plant biotechnology journal · 2026Article
- Improving transient protein expression in agroinfiltrated Nicotiana benthamiana.The New phytologist · 2024Review
- Heterologous expression of influenza haemagglutinin leads to early and transient activation of the unfolded protein response in Nicotiana benthamiana.Plant biotechnology journal · 2024Article
- Optimising expression and extraction of recombinant proteins in plants.Frontiers in plant science · 2022Review
- Co-expression With Replicating Vector Overcoming Competitive Effects Derived by a Companion Protease Inhibitor in Plants.Frontiers in plant science · 2021Article
- A Rationally Designed Bovine IgA Fc Scaffold EnhancesFrontiers in plant science · 2021Article
- Transformation of Amorphadiene Synthase and Antisilencing P19 Genes intoAdvanced pharmaceutical bulletin · 2020Article
- Innovation in plant-based transient protein expression for infectious disease prevention and preparedness.Current opinion in biotechnology · 2020Review
- Enhanced foreign protein accumulation in Nicotiana benthamiana leaves co-infiltrated with a TMV vector and plant cell cycle regulator genes.Transgenic research · 2019Article
- Activity-based proteomics reveals nine target proteases for the recombinant protein-stabilizing inhibitor SlCYS8 in Nicotiana benthamiana.Plant biotechnology journal · 2019Article
- Plant Serine Protease Inhibitors: Biotechnology Application in Agriculture and Molecular Farming.International journal of molecular sciences · 2019Review
- The Rapid Methylation of T-DNAs UponFrontiers in plant science · 2019Article
- Production of Biopharmaceuticals inFrontiers in plant science · 2019Article
- Assessing Carnivorous Plants for the Production of Recombinant Proteins.Frontiers in plant science · 2019Article
- Recombinant protein susceptibility to proteolysis in the plant cell secretory pathway is pH-dependent.Plant biotechnology journal · 2018Article
- Three unrelated protease inhibitors enhance accumulation of pharmaceutical recombinant proteins in Nicotiana benthamiana.Plant biotechnology journal · 2018Article
- Vacuolar targeting of recombinant antibodies in Nicotiana benthamiana.Plant biotechnology journal · 2016Article
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The expression of clinically useful proteins in plants has been bolstered by the development of high-yielding systems for transient protein expression using agroinfiltration. There is a need now to know more about how host plant development and metabolism influence the quantity and quality of recombinant proteins. Endogenous proteolysis is a key determinant of the stability and yield of recombinant proteins in plants. Here we characterised cysteine (C1A) and aspartate (A1) protease profiles in leaves of the widely used expression host Nicotiana benthamiana, in relation with the production of a murine IgG, C5-1, targeted to the cell secretory pathway. Agroinfiltration significantly altered the distribution of C1A and A1 proteases along the leaf age gradient, with a correlation between leaf age and the level of proteolysis in whole-cell and apoplast protein extracts. The co-expression of tomato cystatin SlCYS8, an inhibitor of C1A proteases, alongside C5-1 increased antibody yield by nearly 40% after the usual 6-days incubation period, up to ~3 mg per plant. No positive effect of SlCYS8 was observed in oldest leaves, in line with an increased level of C1A protease activity and a very low expression rate of the inhibitor. By contrast, C5-1 yield was greater by an additional 40% following 8- to 10-days incubations in younger leaves, where high SlCYS8 expression was maintained. These findings confirm that the co-expression of recombinant protease inhibitors is a promising strategy for increasing recombinant protein yields in plants, but that further opportunity exists to improve this approach by addressing the influence of leaf age and proteases of other classes.
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