Rapid expression of COVID-19 proteins by transient expression in tobacco

Lindsay, Penelope L; Ackerman, Amanda; Jian, Yinan; Artz, Oliver; Rosado, Daniele; Skopelitis, Tara; Kitagawa, Munenori; Pedmale, Ullas V.; Jackson, David

doi:10.1101/2020.12.29.424712

Cited by 3 publications

(2 citation statements)

References 22 publications

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“…Over the last decades, plants have been efficiently used as platforms for the rapid and scalable production of functional recombinant proteins [ 1 ]. Plant-based production of recombinant proteins (e.g., vaccines, therapeutics, diagnostic compounds, industrial enzymes) is now a rapidly developing segment of the bioeconomy, with recent data showing how it can contribute to global responses in emergency scenarios [ 2 ], as was the case for the rapid production of SARS-CoV-2 proteins for COVID-19 research by the transient transformation of tobacco plants [ 3 ].…”

Section: Introductionmentioning

confidence: 99%

Enhancing the Production of Hydrolytic Enzymes in Elicited Tamarillo (Solanum betaceum Cav.) Cell Suspension Cultures

Casimiro

Mota

Verı́ssimo

et al. 2023

Plants

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Plant cell suspension cultures are widely used as a tool for analyzing cellular and molecular processes, metabolite synthesis, and differentiation, bypassing the structural complexity of plants. Within the range of approaches used to increase the production of metabolites by plant cells, one of the most recurrent is applying elicitors capable of stimulating metabolic pathways related to defense mechanisms. Previous proteomics analysis of tamarillo cell lines and cell suspension cultures have been used to further characterize and optimize the growth and stress-related metabolite production under in vitro controlled conditions. The main objective of this work was to develop a novel plant-based bioreactor system to produce hydrolytic enzymes using an elicitation approach. Based on effective protocols for tamarillo micropropagation and plant cell suspension culture establishment from induced callus lines, cell growth has been optimized, and enzymatic activity profiles under in vitro controlled conditions characterized. By testing different sucrose concentrations and the effects of two types of biotic elicitors, it was found that 3% (w/v) sucrose concentration in the liquid medium enhanced the production of hydrolytic enzymes. Moreover, casein hydrolysate at 0.5 and 1.5 g/L promoted protein production, whereas yeast extract (0.5 g/L) enhanced glycosidase activity. Meanwhile, chitosan (0.05 and 0.1 g/L) enhanced glycosidases, alkaline phosphates, and protease activities.

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Section: Introductionmentioning

confidence: 99%

Enhancing the Production of Hydrolytic Enzymes in Elicited Tamarillo (Solanum betaceum Cav.) Cell Suspension Cultures

Casimiro

Mota

Verı́ssimo

et al. 2023

Plants

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“…However, as compared to stable transgenic production of recombinant proteins in plants, a transient production system offers a faster and higher level of production (Yamamoto et al 2018). Various industrially important proteins have already been successfully produced via transient expression in plants such as α-amylase (Prado et al 2019), Interleukin-6 (Nausch et al 2012), antibodies (Vézina et al 2009), HIV-1 Nef (Lombardi et al 2009), eVLPs (Sainsbury and Lomonossoff 2008), and even COVID-19 proteins (Lindsay et al 2020). Despite its benefits, the plant expression platform has yet to overcome its major challenges in terms of mass production and profitability (Habibi et al 2017).…”

Section: Introductionmentioning

confidence: 99%

High-level transient production of a protease-resistant mutant form of human basic fibroblast growth factor in <i>Nicotiana benthamiana</i> leaves

Macauyag

Kajiura

Ohashi

et al. 2022

Plant Biotechnology

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The human basic fibroblast growth factor (bFGF) is a protein that plays a pivotal role in cellular processes like cell proliferation and development. As a result, it has become an important component in cell culture systems, with applications in biomedical engineering, cosmetics, and research. Alternative production techniques, such as transient production in plants, are becoming a feasible option as the demand continues to grow. High-level bFGF production was achieved in this study employing an optimized Agrobacterium-mediated transient expression system, which yielded about a 3-fold increase in production over a conventional system. This yield was further doubled at about 185 µg g −1 FW using a mutant protease-resistant version that degraded/aggregated at a three-fold slower rate in leaf crude extracts. To achieve a pure product, a two-step purification technique was applied. The capacity of the pure protease-resistant bFGF (PRbFGF) to stimulate cell proliferation was tested and was found to be comparable to that of E. coli-produced bFGF in HepG2 and CHO-K1 cells. Overall, this study demonstrates a high-level transient production system of functional PRbFGF in N. benthamiana leaves as well as an efficient tag-less purification technique of leaf crude extracts.

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Tamarillo cell suspension-based bioreactors – optimization of a novel system for elicited enzymatic production

Casimiro¹,

Lopes²,

Correia³

et al. 2023

Acta Hortic.

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Rapid expression of COVID-19 proteins by transient expression in tobacco

Cited by 3 publications

References 22 publications

Enhancing the Production of Hydrolytic Enzymes in Elicited Tamarillo (Solanum betaceum Cav.) Cell Suspension Cultures

Enhancing the Production of Hydrolytic Enzymes in Elicited Tamarillo (Solanum betaceum Cav.) Cell Suspension Cultures

High-level transient production of a protease-resistant mutant form of human basic fibroblast growth factor in <i>Nicotiana benthamiana</i> leaves

Tamarillo cell suspension-based bioreactors – optimization of a novel system for elicited enzymatic production

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