High‐yield production of authentic human growth hormone using a plant virus‐based expression system

Gils, Mario; Kandzia, Romy; Marillonnet, Sylvestre; Klimyuk, Victor; Gleba, Yuri

doi:10.1111/j.1467-7652.2005.00154.x

Cited by 115 publications

(91 citation statements)

References 36 publications

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“…In comparison with our CMV system, CMV would be easier in preparation for an inoculum but agroinfiltration would be better in infection efficiency. They also succeeded in high-yield production (10% of soluble protein) of active human growth hormone using the same system (Gils et al 2005). However, in this system, they extracted the expressed protein from the Agrobacteirum-infiltrated areas before the tissues died.…”

Section: Discussionmentioning

confidence: 95%

“…For example, we can mass-produce therapeutic proteins in their active forms in edible plants, enabling the oral-delivery of protein drugs (Arazi et al, 2001;Ma et al, 2005). Currently, heterologous genes can be expressed in plants using either transgenic plants or plant viral vectors (Matsumura et al, 2000;Menassa et al, 2000;Gils et al, 2005). The disadvantages of transgenic plants reside in the system itself; the method is time-consuming and an efficient transformation system is lacking for many plants.…”

Section: Introductionmentioning

confidence: 99%

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Development of Cucumber mosaic virus as a vector modifiable for different host species to produce therapeutic proteins

Matsuo

Hong

Tabayashi

et al. 2006

Planta

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We have developed Cucumber mosaic virus (CMV) as a plant virus vector especially for production of pharmaceutical proteins. The CMV vector is a vector modifiable for different host plants and does not require further engineering steps. CMV contains three genomic RNA molecules (RNAs 1-3) necessary for infectivity. With this system, instead of creating different vector constructs for each plant we use, we take advantage of the formation of pseudrecombinants between two CMV isolates by simply reassembling a vector construct (RNA 2 base) and an RNA molecule containing the host determinant (mostly RNA 3). In this study, the gene for acidic fibroblast growth factor (aFGF), one of the human cytokines, was cloned under the control of the subgenomic promoter for RNA 4A of the CMV-based vector, C2-H1. Infected Nicotiana benthamiana plants produced aFGF at levels up to 5-8% of the total soluble protein. The tobacco-produced aFGF was purified, and its biological activity was confirmed. Using this system, which provides a versatile and viable strategy for production of therapeutic proteins in plants, we also demonstrated a high level of aFGF in Glycine max (soybean) and Arabidopsis thaliana.

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Section: Discussionmentioning

confidence: 95%

Section: Introductionmentioning

confidence: 99%

Development of Cucumber mosaic virus as a vector modifiable for different host species to produce therapeutic proteins

Matsuo

Hong

Tabayashi

et al. 2006

Planta

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“…Further, the chloroplast targeting sequence may act as a translational enhancer, and/or it could be protecting nascent proteins from degradation in the cytosol (Jang et al, 1999). A recent report indicated that accumulation of a human growth hormone protein increased when it was targeted to apoplast, rather than to the chloroplast (Gils et al, 2005). Interestingly, a high level of viral RNAs was present at 18 d.p.i.…”

Section: Discussionmentioning

confidence: 99%

Amplicon-plus targeting technology (APTT) for rapid production of a highly unstable vaccine protein in tobacco plants

et al. 2007

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High-level expression of transgenes is essential for cost-effective production of valuable pharmaceutical proteins in plants. However, transgenic proteins often accumulate in plants at low levels. Low levels of protein accumulation can be caused by many factors including post-transcriptional gene silencing (PTGS) and/or rapid turnover of the transgenic proteins. We have developed an Amplicon-plus Targeting Technology (APTT), by using novel combination of known techniques that appears to overcome both of these factors. By using this technology, we have successfully expressed the highly-labile L1 protein of canine oral papillomavirus (COPV L1) by infecting transgenic tobacco plants expressing a suppressor of post-transcriptional gene silencing (PTGS) with a PVX amplicon carrying a gene encoding L1, and targeting the vaccine protein into the chloroplasts. Further, a scalable ''wound-and-agrospray'' inoculation method has been developed that will permit high-throughput Agrobacterium inoculation of Nicotiana tabacum, and a spray-only method (named ''agrospray'') for use with N. benthamiana to allow large-scale application of this technology. The good yield and short interval from inoculation to harvest characteristic of APTT, combined with the potential for high-throughput achieved by use of the agrospray inoculation protocol, make this system a very promising technology for producing high value recombinant proteins, especially those known to be highly labile, in plants for a wide range of applications including producing vaccines against rapidly evolving pathogens and for the rapid response needed to meet bio-defense emergencies.

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“…24 MagnICON, a TMV-based viral provector system developed by Icon genetics, 25 has shown great promise in rapid production of large quantities of various recombinant proteins. [26][27][28][29][30] Using this system, we have achieved high yield (5-10% TSP) production of PyMSP4/5, which was highly stable at ambient temperature after freeze-drying and was immunogenic following oral administration in mice. 31 Since Mason et al reported the successful expression of HBsAg in transgenic tobacco, which demonstrated the promise of using transgenic plants as low-cost vaccine production systems, 32 the concept of plant-produced vaccine has changed from "edible vaccines" to "heat-stable oral vaccines" or low-cost injectable vaccines.…”

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confidence: 99%

Alternative approaches to vaccine delivery

Wang

Chung

Coppel³

2010

Human Vaccines

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High‐yield production of authentic human growth hormone using a plant virus‐based expression system

Abstract: SummaryWe describe here a high-yield transient expression system for the production of human growth hormone (hGH, or somatotropin)

Cited by 115 publications

References 36 publications

Development of Cucumber mosaic virus as a vector modifiable for different host species to produce therapeutic proteins

Development of Cucumber mosaic virus as a vector modifiable for different host species to produce therapeutic proteins

Amplicon-plus targeting technology (APTT) for rapid production of a highly unstable vaccine protein in tobacco plants

Alternative approaches to vaccine delivery

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