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2.
Molecules ; 26(13)2021 Jun 24.
Artigo em Inglês | MEDLINE | ID: mdl-34202844

RESUMO

The COVID-19 pandemic, as well as the more general global increase in viral diseases, has led researchers to look to the plant kingdom as a potential source for antiviral compounds. Since ancient times, herbal medicines have been extensively applied in the treatment and prevention of various infectious diseases in different traditional systems. The purpose of this review is to highlight the potential antiviral activity of plant compounds as effective and reliable agents against viral infections, especially by viruses from the coronavirus group. Various antiviral mechanisms shown by crude plant extracts and plant-derived bioactive compounds are discussed. The understanding of the action mechanisms of complex plant extract and isolated plant-derived compounds will help pave the way towards the combat of this life-threatening disease. Further, molecular docking studies, in silico analyses of extracted compounds, and future prospects are included. The in vitro production of antiviral chemical compounds from plants using molecular pharming is also considered. Notably, hairy root cultures represent a promising and sustainable way to obtain a range of biologically active compounds that may be applied in the development of novel antiviral agents.


Assuntos
Antivirais/farmacologia , COVID-19/tratamento farmacológico , Extratos Vegetais/farmacologia , Plantas Medicinais/química , SARS-CoV-2/efeitos dos fármacos , Antivirais/química , Antivirais/imunologia , Antivirais/uso terapêutico , Simulação por Computador , Humanos , Agricultura Molecular/métodos , Extratos Vegetais/química , Extratos Vegetais/imunologia , Extratos Vegetais/uso terapêutico , Plantas Medicinais/imunologia , SARS-CoV-2/fisiologia , Replicação Viral/efeitos dos fármacos
3.
Plant Biotechnol J ; 19(10): 1921-1936, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34181810

RESUMO

The fight against infectious diseases often focuses on epidemics and pandemics, which demand urgent resources and command attention from the health authorities and media. However, the vast majority of deaths caused by infectious diseases occur in endemic zones, particularly in developing countries, placing a disproportionate burden on underfunded health systems and often requiring international interventions. The provision of vaccines and other biologics is hampered not only by the high cost and limited scalability of traditional manufacturing platforms based on microbial and animal cells, but also by challenges caused by distribution and storage, particularly in regions without a complete cold chain. In this review article, we consider the potential of molecular farming to address the challenges of endemic and re-emerging diseases, focusing on edible plants for the development of oral drugs. Key recent developments in this field include successful clinical trials based on orally delivered dried leaves of Artemisia annua against malarial parasite strains resistant to artemisinin combination therapy, the ability to produce clinical-grade protein drugs in leaves to treat infectious diseases and the long-term storage of protein drugs in dried leaves at ambient temperatures. Recent FDA approval of the first orally delivered protein drug encapsulated in plant cells to treat peanut allergy has opened the door for the development of affordable oral drugs that can be manufactured and distributed in remote areas without cold storage infrastructure and that eliminate the need for expensive purification steps and sterile delivery by injection.


Assuntos
Artemisia annua , Doenças Transmissíveis , Preparações Farmacêuticas , Animais , Humanos , Agricultura Molecular , Plantas Comestíveis
4.
J Plant Physiol ; 258-259: 153359, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33460995

RESUMO

Recombinant proteins play an important role in many areas of our lives. For example, recombinant enzymes are used in the food and chemical industries and as high-quality proteins for research, diagnostic and therapeutic applications. The production of recombinant proteins is still dominated by expression systems based on microbes and mammalian cells, although the manufacturing of recombinant proteins in plants - known as molecular farming - has been promoted as an alternative, cost-efficient strategy for three decades. Several molecular farming products have reached the market, but the number of success stories has been limited by industrial inertia driven by perceptions of low productivity, the high cost of downstream processing, and regulatory hurdles that create barriers to translation. Here, we discuss the technical and economic factors required for the successful commercialization of molecular farming, and consider potential future directions to enable the broader application of production platforms based on plants.


Assuntos
Agricultura Molecular , Proteínas de Plantas/análise , Plantas Geneticamente Modificadas/química , Proteínas Recombinantes/análise , Biotecnologia
5.
Can J Microbiol ; 67(1): 85-97, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32721220

RESUMO

Agroinfiltration is used to treat plants with modified strains of Agrobacterium tumefaciens for the purpose of transient in planta expression of genes transferred from the bacterium. These genes encode valuable recombinant proteins for therapeutic or industrial applications. Treatment of large quantities of plants for industrial-scale protein production exposes bacteria (harboring genes of interest) to agroinfiltration medium that is devoid of nutrients and carbon sources for prolonged periods of time (possibly upwards of 24 h). Such conditions may negatively influence bacterial viability, infectivity of plant cells, and target protein production. Here, we explored the role of timing in bacterial culture preparation for agroinfiltration using mass spectrometry-based proteomics to define changes in cellular processes. We observed distinct profiles associated with bacterial treatment conditions and exposure timing, including significant changes in proteins involved in pathogenesis, motility, and nutrient acquisition systems as the bacteria adapt to the new environment. These data suggest a progression towards increased cellular remodelling over time. In addition, we described changes in growth- and environment-specific processes over time, underscoring the interconnectivity of pathogenesis and chemotaxis-associated proteins with transport and metabolism. Overall, our results have important implications for the production of transiently expressed target protein products, as prolonged exposure to agroinfiltration medium suggests remodelling of the bacterial proteins towards enhanced infection of plant cells.


Assuntos
Adaptação Fisiológica/efeitos dos fármacos , Inoculantes Agrícolas/efeitos dos fármacos , Agrobacterium tumefaciens/efeitos dos fármacos , Meios de Cultura/farmacologia , Agricultura Molecular , Inoculantes Agrícolas/fisiologia , Agrobacterium tumefaciens/fisiologia , Proteínas de Bactérias/metabolismo , Meios de Cultura/metabolismo , Plantas Geneticamente Modificadas/microbiologia , Proteômica , Proteínas Recombinantes/genética
6.
Can J Microbiol ; 67(1): 75-84, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32846104

RESUMO

The preparation of Agrobacterium tumefaciens cultures with strains encoding proteins intended for therapeutic or industrial purposes is an important activity prior to treatment of plants for transient expression of valuable protein products. The rising demand for biologic products such as these underscores the expansion of molecular pharming and warrants the need to produce transformed plants at an industrial scale. This requires large quantities of A. tumefaciens culture, which is challenging using traditional growth methods (e.g., shake flask). To overcome this limitation, we investigate the use of bioreactors as an alternative to shake flasks to meet production demands. Here, we observe differences in bacterial growth among the tested parameters and define conditions for consistent bacterial culturing between shake flask and bioreactor. Quantitative proteomic profiling of cultures from each growth condition defines unique growth-specific responses in bacterial protein abundance and highlights the functional roles of these proteins, which may influence bacterial processes important for effective agroinfiltration and transformation. Overall, our study establishes and optimizes comparable growth conditions for shake flask versus bioreactors and provides novel insights into fundamental biological processes of A. tumefaciens influenced by such growth conditions.


Assuntos
Agrobacterium tumefaciens/crescimento & desenvolvimento , Agrobacterium tumefaciens/metabolismo , Reatores Biológicos/microbiologia , Agricultura Molecular/métodos , Proteínas de Bactérias/biossíntese , Técnicas de Cultura Celular por Lotes/instrumentação , Técnicas de Cultura Celular por Lotes/métodos , Proteômica
7.
Biotechnol Adv ; 46: 107681, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33326816

RESUMO

The market for biopharmaceuticals is dominated by recombinant proteins and is driven mainly by the development of vaccines and antibodies. Manufacturing predominantly relies on fermentation-based production platforms, which have limited scalability and suffer from high upstream process costs. As an alternative, the production of recombinant proteins in whole plants (plant molecular farming) provides a scalable and cost efficient upstream process because each plant functions as a self-contained bioreactor, avoiding costs associated with single-use devices and cleaning-in-place. Despite many proof-of-concept studies and the approval of a few products as medical devices, the only approved pharmaceutical proteins manufactured in whole plants have been authorized under emergency protocols. The absence of approvals under standard clinical development pathways in part reflects the lack of standardized process equipment and unit operations, leading to industry inertia based on familiarity with fermenter systems. Here we discuss the upstream production steps of plant molecular farming by transient expression in intact plants, including seeding, plant cultivation, infiltration with Agrobacterium tumefaciens, post-infiltration incubation, and harvesting. We focus on cultivation techniques because they strongly affect the subsequent steps and overall process design. We compare the benefits and drawbacks of open field, greenhouse and vertical farm strategies in terms of upfront investment costs, batch reproducibility, and decoupling from environmental impacts. We consider process automation, monitoring and adaptive process design in the context of Industry 4.0, which can boost process efficiency and batch-to-batch uniformity to improve regulatory compliance. Finally, we discuss the costs-benefit aspects of the different cultivation systems.


Assuntos
Produtos Biológicos , Agricultura Molecular , Plantas , Plantas Geneticamente Modificadas/genética , Proteínas Recombinantes/genética , Reprodutibilidade dos Testes
8.
Genes (Basel) ; 11(9)2020 08 19.
Artigo em Inglês | MEDLINE | ID: mdl-32825160

RESUMO

We set out to assess the NIH/3T3 cell proliferation activity of Arabidopsis oil body-expressed recombinant oleosin-hEGF-hEGF protein. Normally, human epidermal growth factor (hEGF) is purified through complex process, however, oleosin fusion technology provides an inexpensive and scalable platform for its purification. Under a phaseolin promoter, we concatenated oleosin gene to double hEGF (hEGF-hEGF) with plant-preferred codons in the expression vectors and the construct was transformed into Arabidopsis thaliana (Arabidopsis). The transgenic Arabidopsis was validated by RT-PCR and the content of recombinant protein oleosin-hEGF-hEGF was quantified by western blot. Subsequently, the proliferation assay and transdermal absorption were determined by MTT method and immunohistochemical staining, respectively. First, the expression level of hEGF was recorded to be 14.83-ng/µL oil body and due to smaller size transgenic oil bodies expressing the recombinant oleosin-hEGF-hEGF, they were more skin permeable than those of control. Second, via the staining intensity of transgenic oil bodies was greater than EGF at all time points via immunohistochemical staining in transdermal absorption process. Lastly, activity assays of oil bodies expressed oleosin-hEGF-hEGF indicated that they stimulated the NIH/3T3 cell proliferation activity. Our results revealed oil-body-expressed oleosin-hEGF-hEGF was potential new material having implications in the field of medicine.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Fator de Crescimento Epidérmico/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Proteínas Recombinantes/metabolismo , Pele/metabolismo , Animais , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Proteínas de Arabidopsis/química , Proteínas de Arabidopsis/genética , Proliferação de Células , Fator de Crescimento Epidérmico/química , Fator de Crescimento Epidérmico/genética , Humanos , Camundongos , Agricultura Molecular , Células NIH 3T3 , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/crescimento & desenvolvimento , Regiões Promotoras Genéticas , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Cicatrização
9.
J Biotechnol ; 322: 10-20, 2020 Oct 10.
Artigo em Inglês | MEDLINE | ID: mdl-32659239

RESUMO

One of the milestones of vaccinology is the depletion of the global impact of Poliomyelitis. The current vaccines to deal with Polio comprise the Sabin and Salk formulations. The main limitation of the former is the use of attenuated viruses that can revert into pathogenic forms, whereas the latter is more expensive and induces no protection in the intestinal tract; the site of virus replication. Genetically engineered plants cope with such limitations. In addition, they offer a low-cost alternative for production, storage and delivery of vaccines. This technology has been narrowly applied in the development of Polio vaccines. Herein, we explored the ability of tobacco cells to express the immunogenic VP1, VP2, VP3, and VP4 Polio antigens, which are relevant for vaccine development. Evidence on the expression of the plant-made Polio VPs is presented and an immunogenicity assessment proved their capacity to induce local and systemic humoral responses when administered by subcutaneous and oral routes. The plant-made VPs will be useful in the development of low-cost vaccine formulations able to induce effective mucosal immunity without the risks associated to the use of attenuated viruses; therefore there is a potential for this technology to contribute toward Polio eradication.


Assuntos
Proteínas do Capsídeo , Vacina Antipólio Oral , Poliovirus , Tabaco/genética , Vacinas de Subunidades , Animais , Anticorpos Antivirais/análise , Anticorpos Antivirais/sangue , Antígenos Virais/genética , Antígenos Virais/imunologia , Antígenos Virais/metabolismo , Proteínas do Capsídeo/genética , Proteínas do Capsídeo/imunologia , Proteínas do Capsídeo/metabolismo , Fezes/química , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Agricultura Molecular , Plantas Geneticamente Modificadas/genética , Poliomielite/prevenção & controle , Poliomielite/virologia , Poliovirus/genética , Poliovirus/imunologia , Vacina Antipólio Oral/genética , Vacina Antipólio Oral/imunologia , Vacina Antipólio Oral/metabolismo , Vacinas de Subunidades/genética , Vacinas de Subunidades/imunologia , Vacinas de Subunidades/metabolismo
10.
Expert Opin Biol Ther ; 20(6): 545-548, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32250170

RESUMO

Given the dramatic impact of the COVID-19 pandemic, it is imperative to divulge all the available technologies with the potential to fight against this virus. Plant biotechnology offers potential solutions to this pandemic through the development of low-cost vaccines and antibodies useful for therapy, prophylaxis, and diagnosis. The technology to produce plant-made biopharmaceuticals is already established; two examples of these are: a therapeutic enzyme that has entered the market and the influenza vaccines that are currently under clinical trials with encouraging results. Thus far, some companies have started developing anti-COVID-19 antibodies and vaccines. In particular, plant-made antibodies might be timely produced and approved for human use in the short term, while the development of vaccines will take longer time (clinical evaluations could be concluded by the end of 2021); nonetheless, the candidates obtained will be valuable tools for future outbreaks. The key aspects that will define the exploitation of this technology in the fight against COVID-19 are discussed.


Assuntos
Produtos Biológicos/uso terapêutico , Infecções por Coronavirus/tratamento farmacológico , Agricultura Molecular , Plantas Geneticamente Modificadas , Pneumonia Viral/tratamento farmacológico , Anticorpos Antivirais/biossíntese , Betacoronavirus , Biotecnologia , COVID-19 , Vacinas contra COVID-19 , Infecções por Coronavirus/prevenção & controle , Humanos , Pandemias , SARS-CoV-2 , Vacinas Virais/biossíntese
11.
PLoS One ; 15(3): e0229952, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32196508

RESUMO

Plant molecular farming (PMF) is a convenient and cost-effective way to produce high-value recombinant proteins that can be used in the production of a range of health products, from pharmaceutical therapeutics to cosmetic products. New plant breeding techniques (NPBTs) provide a means to enhance PMF systems more quickly and with greater precision than ever before. However, the feasibility, regulatory standing and social acceptability of both PMF and NPBTs are in question. This paper explores the perceptions of key stakeholders on two European Union (EU) Horizon 2020 programmes-Pharma-Factory and Newcotiana-towards the barriers and facilitators of PMF and NPBTs in Europe. One-on-one qualitative interviews were undertaken with N = 20 individuals involved in one or both of the two projects at 16 institutions in seven countries (Belgium, France, Germany, Italy, Israel, Spain and the UK). The findings indicate that the current EU regulatory environment and the perception of the public towards biotechnology are seen as the main barriers to scaling-up PMF and NPBTs. Competition from existing systems and the lack of plant-specific regulations likewise present challenges for PMF developing beyond its current niche. However, respondents felt that the communication of the benefits and purpose of NPBT PMF could provide a platform for improving the social acceptance of genetic modification. The importance of the media in this process was highlighted. This article also uses the multi-level perspective to explore the ways in which NPBTs are being legitimated by interested parties and the systemic factors that have shaped and are continuing to shape the development of PMF in Europe.


Assuntos
Agricultura Molecular , Melhoramento Vegetal , Plantas/genética , Tabaco/crescimento & desenvolvimento , Biotecnologia , União Europeia , Edição de Genes , Engenharia Genética/tendências , Humanos , Desenvolvimento Vegetal/genética , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/crescimento & desenvolvimento
12.
Mol Biotechnol ; 62(4): 240-251, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32108286

RESUMO

In the past decade, interest in the production of recombinant pharmaceutical proteins in plants has tremendously progressed because plants do not harbor mammalian viruses, are economically competitive, easily scalable, and capable of carrying out complex post-translational modifications required for recombinant pharmaceutical proteins. Mucuna bracteata is an essential perennial cover crop species widely planted as an underground cover in oil palm and rubber plantations. As a legume, they have high biomass, thrive in its habitat, and can fix nitrogen. Thus, M. bracteata is a cost-efficient crop that shows ideal characteristics as a platform for mass production of recombinant protein. In this study, we established a new platform for the transient production of a recombinant protein in M. bracteata via vacuum-assisted agro-infiltration. Five-week-old M. bracteata plants were vacuum infiltrated with Agrobacterium tumefaciens harboring a plasmid that encodes for an anti-toxoplasma immunoglobulin (IgG) under different parameters, including trifoliate leaf positional effects, days to harvest post-infiltration, and the Agrobacterium strain used. Our results showed that vacuum infiltration of M. bracteata plant with A. tumefaciens strain GV3101 produced the highest concentration of heterologous protein in its bottom trifoliate leaf at 2 days post-infiltration. The purified anti-toxoplasma IgG was then analyzed using Western blot and ELISA. It was demonstrated that, while structural heterogeneity existed in the purified anti-toxoplasma IgG from M. bracteata, its transient expression level was two-fold higher than the model platform, Nicotiana benthamiana. This study has laid the foundation towards establishing M. bracteata as a potential platform for the production of recombinant pharmaceutical protein.


Assuntos
Imunoglobulina G/biossíntese , Agricultura Molecular/métodos , Mucuna/genética , Agrobacterium tumefaciens/genética , Expressão Gênica , Técnicas de Transferência de Genes/instrumentação , Imunoglobulina G/genética , Imunoglobulina G/imunologia , Imunoglobulina G/isolamento & purificação , Mucuna/metabolismo , Plantas Geneticamente Modificadas/genética , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/farmacologia , Fatores de Tempo , Toxoplasma/imunologia , Transformação Bacteriana
13.
Biotechnol Adv ; 40: 107519, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31954848

RESUMO

Molecular farming can be defined as the use of plants to produce recombinant protein products. The technology is now >30 years old. The early promise of molecular farming was based on three perceived advantages: the low costs of growing plants, the immense scalability of agricultural production, and the inherent safety of plants as hosts for the production of pharmaceuticals. This resulted in a glut of research publications in which diverse proteins were expressed in equally diverse plant-based systems, and numerous companies were founded hoping to commercialize the new technology. There was a moderate degree of success for companies producing non-pharmaceutical proteins, but in the pharmaceutical sector the anticipation raised by promising early research was soon met by the cold hard reality of industrial pragmatism. Plants did not have a track record of success in pharmaceutical protein manufacturing, lacked a regulatory framework, and did not perform as well as established industry platforms. Negative attitudes towards genetically modified plants added to the mix. By the early 2000s, major industry players started to lose interest and pharmaceutical molecular farming fell from a peak of expectation into a trough of disillusionment, just as predicted by the Gartner hype cycle. But many of the pioneers of molecular farming have refocused their activities and have worked to address the limitations that hampered the first generation of technologies. The field has now consolidated around a smaller number of better-characterized platforms and has started to develop standardized methods and best practices, mirroring the evolution of more mature industry sectors. Likewise, attention has turned from proof-of-principle studies to realistic techno-economic modeling to capture significant niche markets, replicating the success of the industrial molecular farming sector. Here we argue that these recent developments signify that pharmaceutical molecular farming is now climbing the slope of enlightenment and will soon emerge as a mature technology.


Assuntos
Agricultura Molecular , Plantas Geneticamente Modificadas , Proteínas Recombinantes
14.
Curr Opin Biotechnol ; 61: 15-20, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31593785

RESUMO

The production of biopharmaceuticals in plant-based systems had faced several challenges that hampered broader adoption of this technology. In recent years, various plant production hosts have been improved by genetic engineering approaches to overcome obstacles with regard to post-translational modifications and integrity of target proteins. Together with optimized extraction and purification processes, those advances have put plant molecular farming in a more competitive position compared to established production systems. Certain biopharmaceuticals can be derived from plant systems with unique desired properties, qualifying them as biobetters.


Assuntos
Produtos Biológicos , Agricultura Molecular , Engenharia Genética , Plantas Geneticamente Modificadas/genética , Processamento de Proteína Pós-Traducional , Proteínas Recombinantes
15.
Artigo em Inglês | MEDLINE | ID: mdl-31486296

RESUMO

The use of plants for the production of virus-like nanoparticles (VNPs) dates back to separating natural empty capsids of plant viruses from whole virions nearly 70 years ago, through to the present use of transgenic plants or recombinant Agrobacterium tumefaciens and/or plant virus-derived vectors for the transient expression of engineered viral or other structural proteins in plants-a production system also known as molecular farming. Plant production of heterologous proteins has major advantages in terms of convenience-whole plants are generally used, and processes do not need to be sterile-and cost, as bulk biomass production is significantly cheaper than by any other method. Plant-made VNPs in current use for nanotechnology include whole virions and naturally occurring empty capsids of plant viruses, and particles made by reassembly of coat protein (CP) purified from virions or by recombinant expression. Engineered VNP-forming animal or human virus CPs expressed in plants include L1 protein from human papillomaviruses, human norovirus CP, hepatitis B surface and core antigens, influenza virus HA protein and HIV Gag polyprotein forming large enveloped particles by budding, orbi- and rotavirus particles that require assembly of four co-expressed proteins, and polio- and foot and mouth disease viruses which require proteolytic processing of a polyprotein precursor to form 4-component VNPs. Both plant and animal virus-derived plant-made VNPs can be used for surface and internal display of heterologous peptides or even whole proteins. A significant recent development has been the production of pseudovirions in plants, comprising plant or animal virus CPs and RNA or DNA pseudogenomes that can be used to deliver nucleic acid payloads into cultured cells or specific tissues or tumors in whole animals. This article is characterized under: Biology-Inspired Nanomaterials > Protein and Virus-Based Structures Therapeutic Approaches and Drug Discovery > Emerging Technologies Diagnostic Tools > in vivo Nanodiagnostics and Imaging.


Assuntos
Agricultura Molecular , Nanopartículas , Vírion , Animais , Indicadores e Reagentes , Plantas Geneticamente Modificadas , Vacinas
16.
Curr Opin Biotechnol ; 61: 89-95, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31786432

RESUMO

Local manufacturing of protein-based vaccines and therapies in Africa is limited and contributes to a trade deficit, security of supply concerns and poor access to biopharmaceuticals by the poor. Plant molecular farming is a potential technology solution that has received growing adoption by African scientists attracted by the potential for the competitive cost of goods, safety and efficacy. Plant-made pharmaceutical technologies for veterinary and human vaccination and treatment of non-communicable and infectious diseases are available at different stages of development in Africa. There is also growth in the translation of these technologies to commercial operations. Africa is poised to benefit from the real-world impact of molecular farming in the next few years.


Assuntos
Doenças Transmissíveis , Vacinas , África , Humanos , Agricultura Molecular , Vacinação
17.
Curr Opin Biotechnol ; 61: 60-65, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31765962

RESUMO

Molecular farming increasingly uses the tobacco relative Nicotiana benthamiana for production of recombinant proteins through transient expression. Several proteins are produced efficiently with this expression platform, but yields for other proteins are often very low. These low yields are frequently due to endogenous proteases. The latest genome annotations indicate that N. benthamiana encodes for at least 1243 putative proteases that probably act redundantly and consecutively on substrates in different subcellular compartments. Here, we discuss the N. benthamiana protease repertoire that may affect recombinant protein production and recent advances in protease depletion strategies to increase recombinant protein production in N. benthamiana.


Assuntos
Agricultura Molecular , Tabaco/genética , Endopeptidases , Peptídeo Hidrolases , Plantas Geneticamente Modificadas , Proteínas Recombinantes
18.
Mol Biotechnol ; 62(2): 79-90, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31758488

RESUMO

Vaccines are biological preparations that improve immunity to particular diseases and form an important innovation of 19th century research. It contains a protein that resembles a disease-causing microorganism and is often made from weak or killed forms of the microbe. Vaccines are agents that stimulate the body's immune system to recognize the antigen. Now, a new form of vaccine was introduced which will have the power to mask the risk side of conventional vaccines. This type of vaccine was produced from plants which are genetically modified. In the production of edible vaccines, the gene-encoding bacterial or viral disease-causing agent can be incorporated in plants without losing its immunogenic property. The main mechanism of action of edible vaccines is to activate the systemic and mucosal immunity responses against a foreign disease-causing organism. Edible vaccines can be produced by incorporating transgene in to the selected plant cell. At present edible vaccine are developed for veterinary and human use. But the main challenge faced by edible vaccine is its acceptance by the population so that it is necessary to make aware the society about its use and benefits. When compared to other traditional vaccines, edible vaccines are cost effective, efficient and safe. It promises a better prevention option from diseases.


Assuntos
Produtos Biológicos/imunologia , Imunidade nas Mucosas/efeitos dos fármacos , Organismos Geneticamente Modificados/imunologia , Plantas Geneticamente Modificadas/imunologia , Vacinas de Plantas Comestíveis/imunologia , Administração Oral , Agrobacterium/genética , Agrobacterium/imunologia , Animais , Biolística/métodos , Clorófitas/genética , Clorófitas/imunologia , Técnicas de Transferência de Genes , Humanos , Insetos/genética , Insetos/imunologia , Lactobacillales/genética , Lactobacillales/imunologia , Agricultura Molecular , Vírus de Plantas/genética , Vírus de Plantas/imunologia , Leveduras/genética , Leveduras/imunologia
19.
Curr Opin Biotechnol ; 61: 53-59, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31751895

RESUMO

Interest in applications and benefits that Molecular Pharming might offer to Low and Middle Income Countries has always been a potent driver for the research discipline, and a major reason why many scientists entered the field. Although enthusiasm remains high, the reality is that such a game-changing innovation would always take longer than traditional uptake of new technology in developed countries, and be complicated by external factors beyond technical feasibility. Excitingly, signs of increasing interest by LMICS in Molecular Pharming are now emerging. Here, three case studies from Thailand, South Africa and Brazil are used to identify some of the key issues when a new investment into Molecular Pharming manufacturing capacity is under consideration. At present, academic research is not necessarily addressing these issues. Only by understanding the concerns, can members of the academic community contribute to helping the development of Molecular Pharming for LMICs by focusing their research efforts appropriately.


Assuntos
Países em Desenvolvimento , Agricultura Molecular , Comércio
20.
Appl Microbiol Biotechnol ; 103(23-24): 9479-9491, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31701198

RESUMO

The successful production of recombinant enzymes by tobacco transplastomic plants must maintain compatibility of the heterologous enzyme with chloroplast metabolism and its long-time enzyme stability. Based on previous reports, it has been taken for granted that following biolistic-transformation, homoplasticity could be obtained from the initially heteroplastic state following successive rounds of selection in the presence of the selection agent. However, several studies indicated that this procedure does not always ensure the complete elimination of unmodified wild-type plastomes. The present study demonstrates that CelK1 transplastomic plants, which were photosyntetically as active as untransformed ones, remain heteroplastomic even after repeated selection steps and that this state does not impair the relatively high-level production of the recombinant enzyme. In fact, even in the heteroplastomic state, the recombinant protein represented about 6% of the total soluble proteins (TSP). Moreover, our data also show that, while the recombinant endoglucanase undergoes phosphorylation, this post-translation modification does not have any significant impact on the enzymatic activity. Biomass storage might be required whenever the enzyme extraction process could not be performed immediately following the harvest of tobacco mature plants. In this respect, we have observed that enzyme activity in the detached leaves stored at 4 °C is maintained up to 20 weeks without significant loss of activity. These findings may have major implications in the future of chloroplast genetic engineering-based molecular farming to produce industrial enzymes in transplastomic plants.


Assuntos
Celulase/biossíntese , Cloroplastos/genética , Agricultura Molecular , Tabaco/enzimologia , Tabaco/genética , Celulase/genética , Engenharia Genética , Microbiologia Industrial/métodos , Folhas de Planta/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Proteoma , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/genética
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