High‐level production of human interleukin‐10 fusions in tobacco cell suspension cultures

Interleukin-11 (IL-11) is a cytokine that plays a key regulatory role in the immune system. Recombinant human IL-11 (rhIL-11) exerts a preventative effect against apoptotic cell death and inhibits preadipocyte differentiation. IL-11 also is used to stimulate the bone marrow to produce platelets in order to prevent low platelets that may be caused by chemotherapy. Unfortunately, the high production cost of IL-11 associated. In this study, we investigated the feasibility of transgenic plants for the cost-effective production of rhIL-11. Production of rhIL-11 proteins in whole-plant expression system will be more economical when compared to the current E. coli based expression system. The human rhIL-11 gene was codon optimized to maximize plant host system expression. IL-11 expression vector under the control of a constitutive cauliflower mosaic virus 35S (CaMV 35S) promoter was introduced into tobacco by Agrobacteriummediated transformation. The 5′-leader sequence (called Ω) of tobacco mosaic virus (TMV) as a translational enhancer was added to construct. Transgenic tobacco plants expressing various levels of rhIL-11 protein were generated. Western blotting of the stably transformed lines demonstrated accumulation of the appropriately sized rhIL-11 protein in leaves. This research demonstrated the efficacy of using tobacco as an expression system for the production of rhIL-11.

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“…Also, the feasibility of transgenic plants for the production of rhIL-4 was demonstrated (Kaldi et al 2013).…”

Section: Introductionmentioning

confidence: 99%

Production of Recombinant Human Interleukin-11 (IL-11) in Transgenic Tobacco (Nicotiana tabacum) Plants

2016

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“…As the demand for safe and inexpensive means for mass production of recombinant pharmaceutical proteins increases, the production of complex mammalian proteins in transformed plants or plant suspension cells, such as HIV-neutralizing antibody 2G12B expressed in tobacco and rice Sack et al, 2015;Vamvaka et al, 2016), human interleukin-10 fusions expressed in tobacco cell suspension cultures (Kaldis et al, 2013), is desirable. Because plants are suitable for the large scale production of pharmaceutical proteins, and the expressed proteins are functional and almost indistinguishable from their mammalian counterparts, they will be available in significantly larger amounts, and a cost-effective basis (Sharma and Sharma, 2009).…”

Section: Discussionmentioning

confidence: 99%

Expression of recombinant human anti-TNF-α scFv-Fc in Arabidopsis thaliana seeds

Yao¹,

Ai²,

Dong³

et al. 2016

Genet. Mol. Res.

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ABSTRACT. Recombinant human anti-tumor necrosis factor (TNF)-α scFv-Fc was expressed in TKO mutant Arabidopsis thaliana seeds using plant-specific codons. Immunoblotting using a human IgG1 antibody detected the expression of anti-TNF-α proteins in plants. Results from qRT-PCR analysis demonstrated that the time of harvest significantly affected the protein yield and quality. Our results indicate that the Phaseolus vulgaris β-phaseolin promoter directed anti-TNF-α scFvFc expression in A. thaliana seeds, with a maximum yield obtained at 20-days of development. Although the yield of anti-TNF-α scFvFc protein was not very high, accumulation of recombinant proteins in seeds is an attractive and simple method that can be used to purify biologically active anti-TNF-α scFv-Fc.

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“…В экспоненциальной фазе роста для удвоения клеток не-обходимо 2-3 суток, один цикл культивирования клеток BY-2 табака, например, составляет 1-2 недели, в то время как на получение растений уходит несколько месяцев (Kaldis et al, 2013).…”

Section: суспензионные клеточные культурыunclassified

Plant expression systems for production of recombinant pharmaceutically important proteins

Дейнеко¹,

Загорская²

2017

Vestn. VOGiS

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The market of pharmaceutically valuable proteins is the fastest growing segment of the economy. Most biopharmaceuticals have been obtained in mammalian and microorganism cells, but both systems have a num ber of disadvantages. Plant cells combine the advan tages of the eukaryotic system of protein production and the simplicity and cheapness of the bacterial, and the use of plants for the production of recombinant proteins is an economically important and promising direction. The advantage of plant systems is the lower cost of cell cultivation. They are free from unwanted components, such as bacterial endotoxins, hyperglyco sylated proteins produced by yeast, animal and human pathogens in cell cultures of transgenic animals. In ad dition, plants are higher eukaryotes, and therefore full value folding and the formation of multimeric protein complexes occur in their cells, as well as a significant portion of posttranslational modifications similar to those in mammalian cells. The currently developed plant expression systems for recombinant proteins are extremely diverse and number more than 100 different technologies based on different plant species, gene transfer methods, expression strategies, methods for the subsequent extraction of the target protein, etc. This is nuclear and plastid transformation, transient and stable expression during transformation using agro bacterial transport, bombardment or electroporation, cultivation of whole terrestrial or aquatic plants, plant tissues or suspension cell cultures as expression sys tems. The review examines the current state of research in the use of plant expression systems for the produc tion of recombinant proteins for pharmaceuticals. The emphasis was placed on the advantages of plant cell cultures in comparison with other expression systems. Specific examples discuss promising plant systems for the production of recombinant proteins, such as trans plastomic plants, moss and aquatic plant cultures, as well as suspension cultures of cells of higher plants. The current state of the market for recombinant proteins obtained using plant expression systems is considered. The prospects of creating plant ("edible") vaccines based on genetically modified plants are discussed.Key words: expression systems; transgenic plants; bio producers; recombinant proteins; plant vaccines.Рынок фармацевтически ценных белков -наиболее быстро разви вающийся сегмент экономики. Большая часть биофармацевтиков получена в клетках млекопитающих и микроорганизмов, однако обе системы обладают рядом недостатков. Растительные клетки сочетают в себе достоинства эукариотической системы наработки белка и простоту и дешевизну бактериальной. Использование рас тений для получения рекомбинантных белков -экономически зна чимое и перспективное направление. Преимуществом раститель ных систем является более низкая стоимость культивирования клеток. Они свободны от нежелательных компонентов, таких как эндотоксины бактерий, гипергликозилированные белки, продуци руемые дрожжами, патогены животных и человека в кле...

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High‐level production of human interleukin‐10 fusions in tobacco cell suspension cultures

Cited by 61 publications

References 53 publications

Production of Recombinant Human Interleukin-11 (IL-11) in Transgenic Tobacco (Nicotiana tabacum) Plants

Production of Recombinant Human Interleukin-11 (IL-11) in Transgenic Tobacco (Nicotiana tabacum) Plants

Expression of recombinant human anti-TNF-α scFv-Fc in Arabidopsis thaliana seeds

Plant expression systems for production of recombinant pharmaceutically important proteins

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