Plant-based production of biopharmaceuticals

Fischer, Rainer; Stöger, Eva; Schillberg, Stefan; Christou, Paul; Twyman, Richard M.

doi:10.1016/j.pbi.2004.01.007

Cited by 570 publications

(376 citation statements)

References 61 publications

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“…Technical advances and limitations of plant-based protein production systems have been recently detailed in (Fischer et al, 2004). We showed here a high yield (2.3% TSP) obtained with a cytosolic protein-expression system.…”

Section: Biotechnological Applicationsmentioning

confidence: 77%

Controlled Expression of Recombinant Proteins in Physcomitrella patens by a Conditional Heat-shock Promoter: a Tool for Plant Research and Biotechnology

et al. 2005

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The ability to express tightly controlled amounts of endogenous and recombinant proteins in plant cells is an essential tool for research and biotechnology. Here, the inducibility of the soybean heat-shock Gmhsp17.3B promoter was addressed in the moss Physcomitrella patens, using b-glucuronidase (GUS) and an F-actin marker (GFP-talin) as reporter proteins. In stably transformed moss lines, Gmhsp17.3B-driven GUS expression was extremely low at 25°C. In contrast, a short non-damaging heat-treatment at 38°C rapidly induced reporter expression over three orders of magnitude, enabling GUS accumulation and the labelling of F-actin cytoskeleton in all cell types and tissues. Induction levels were tightly proportional to the temperature and duration of the heat treatment, allowing fine-tuning of protein expression. Repeated heating/cooling cycles led to the massive GUS accumulation, up to 2.3% of the total soluble proteins. The anti-inflammatory drug acetyl salicylic acid (ASA) and the membrane-fluidiser benzyl alcohol (BA) also induced GUS expression at 25°C, allowing the production of recombinant proteins without heat-treatment. The Gmhsp17.3B promoter thus provides a reliable versatile conditional promoter for the controlled expression of recombinant proteins in the moss P. patens.Abbreviations: ASA, acetyl salicylic acid; BA, benzyl alcohol; GFP, green fluorescent protein; GT, GFPtalin; GUS, b-glucuronidase; Fv/Fm, the maximal photochemical efficiency of PSII; MU, 4-methylumbelliferone; MUG, 4-methylumbelliferyl-b-D-glucuronide; PSII, photosystem II; SA, specific activity; sHSP, small heat-shock protein; TSP, total soluble proteins; Ubi, ubiquitin; X-Gluc, 5-bromo-4-chloro-3-indolyl-b-D-glucuronide

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Section: Biotechnological Applicationsmentioning

confidence: 77%

Controlled Expression of Recombinant Proteins in Physcomitrella patens by a Conditional Heat-shock Promoter: a Tool for Plant Research and Biotechnology

et al. 2005

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“…However, these systems have several disadvantages in terms of cost, stability, safety, etc (Fischer et al, 2004). Plants offer an alternative to microbial fermentation and animal cell cultures and are now gaining widespread acceptance as a general platform for the large-scale production of recombinant proteins (Streatfi eld, 2007).…”

Section: Introductionmentioning

confidence: 99%

“…Plants offer an alternative to microbial fermentation and animal cell cultures and are now gaining widespread acceptance as a general platform for the large-scale production of recombinant proteins (Streatfi eld, 2007). Plants are the most economical producers of biomass and allow for cost-effective production of recombinant proteins on an agricultural scale (Fischer et al, 2004;Berberich et al, 2005). They avoid animal or microbial cell-culture contaminants, such as mammalian pathogens or bacterial components (Fischer et al, 2004;Ma et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

“…Plants are the most economical producers of biomass and allow for cost-effective production of recombinant proteins on an agricultural scale (Fischer et al, 2004;Berberich et al, 2005). They avoid animal or microbial cell-culture contaminants, such as mammalian pathogens or bacterial components (Fischer et al, 2004;Ma et al, 2003). They are also capable of performing complex post-translational modifi cations, most importantly glycosylation, which is often crucial to the biological activity and stability of human therapeutic proteins (Ma et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

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High-level expression of modified gene encoding human adiponectin in transgenic rice

2011

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Adiponectin is a polypeptide specifi cally secreted from human adipocytes, and its defi ciency is closely linked to increased obesity and type II diabetes. There is an urgent demand for large-scale production of human adiponectin for pharmaceutical applications. Here, we report that we have successfully obtained a high-level of expression of modifi ed genes encoding human adiponectin in transgenic rice. The 735 bp cDNA of the native human sequence was adopted to rice codon usage, fused to the translation initiation sequence in the N terminus and to the KDEL signal sequence in the C terminus. An amplifi cation promoting sequence acting as an enhancer of transcription was also introduced to enhance gene expression. The presence of the transgene and mRNA transcripts was confi rmed by PCR, Southern blot and RT-PCR. Western blot analysis revealed that a protein of approximately 30 kDa was produced in rice leaves. ELISA analysis was used to determine the amount of recombinant adiponectin in transformants with the modifi ed gene in up to 0.32% of total soluble leaf protein. Our results establish the feasibility of high-level expression of recombinant human adiponectin in transgenic rice.

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“…Following the first report of successful production of a mouse monoclonal antibody in a plant (Hiatt et al 1989), many other therapeutic proteins have been expressed in transgenic plants. In general, four approaches have been considered for developing molecular farming system in crop species (Fischer et al 2004): nuclear transformation, plastid (chloroplast) transformation, virus-mediated transient transformation, and stable transformation of hydroponically-grown plant species.…”

Section: Introductionmentioning

confidence: 99%

Production of recombinant single chain antibodies (scFv) in vegetatively reproductive Kalanchoe pinnata by in planta transformation

et al. 2009

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We developed an asexual reproductive plant, Kalanchoe pinnata, as a new bioreactor for plant-based molecular farming using a newly developed transformation method. Leaf crenate margins were pin-pricked to infect the plant with the Agrobacterium strain LBA4404 and vacuum infiltration was also applied to introduce the target gene into the plants. Subsequently, the young mother leaf produced new clones at the leaf crenate margins without the need for time-and labor-consuming tissue culture procedures. The average transformation rates were approximately 77 and 84% for pin-prickling and vacuum-infiltration methods, respectively. To functionally characterize an introduced target protein, a nucleic acid hydrolyzing recombinant 3D8 scFv was selected and the plant based 3D8 scFv proteins were purified and analyzed. Based on abzyme analysis, the purified protein expressed with this system had catalytic activity and exhibited all of properties of the protein produced in an E. coli system. This result suggested that vegetatively reproductive K. pinnata can be a novel and potent bioreactor for bio-pharmaceutical proteins.

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Plant-based production of biopharmaceuticals

Cited by 570 publications

References 61 publications

Controlled Expression of Recombinant Proteins in Physcomitrella patens by a Conditional Heat-shock Promoter: a Tool for Plant Research and Biotechnology

Controlled Expression of Recombinant Proteins in Physcomitrella patens by a Conditional Heat-shock Promoter: a Tool for Plant Research and Biotechnology

High-level expression of modified gene encoding human adiponectin in transgenic rice

Production of recombinant single chain antibodies (scFv) in vegetatively reproductive Kalanchoe pinnata by in planta transformation

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