Prevention and treatment of autoimmune diseases with plant virus nanoparticles

Plant viruses are natural, self-assembling nanostructures with versatile and genetically programmable shells, making them useful in diverse applications ranging from the development of new materials to diagnostics and therapeutics. Here, we describe the design and synthesis of plant virus nanoparticles displaying peptides associated with two different autoimmune diseases. Using animal models, we show that the recombinant nanoparticles can prevent autoimmune diabetes and ameliorate rheumatoid arthritis. In both cases, this effect is based on a strictly peptide-related mechanism in which the virus nanoparticle acts both as a peptide scaffold and as an adjuvant, showing an overlapping mechanism of action. This successful preclinical testing could pave the way for the development of plant viruses for the clinical treatment of human autoimmune diseases

Edible plants for oral delivery of biopharmaceuticals

Molecular farming is the use of plants for the production of high value recombinant proteins. Over the last 25\ua0years, molecular farming has achieved the inexpensive, scalable and safe production of pharmaceutical proteins using a range of strategies. One of the most promising approaches is the use of edible plant organs expressing biopharmaceuticals for direct oral delivery. This approach has proven to be efficacious in several clinical vaccination and tolerance induction trials as well as multiple preclinical studies for disease prevention. The production of oral biopharmaceuticals in edible plant tissues could revolutionize the pharmaceutical industry by reducing the cost of production systems based on fermentation, and also eliminating expensive downstream purification, cold storage and transportation costs. This review considers the unique features that make plants ideal as platforms for the oral delivery of protein-based therapeutics and describes recent developments in the production of plant derived biopharmaceuticals for oral administration

Comparative Evaluation of Recombinant Protein Production in Different Biofactories: The Green Perspective

In recent years, the production of recombinant pharmaceutical proteins in heterologous systems has increased significantly. Most applications involve complex proteins and glycoproteins that are difficult to produce, thus promoting the development and improvement of a wide range of production platforms. No individual system is optimal for the production of all recombinant proteins, so the diversity of platforms based on plants offers a significant advantage. Here, we discuss the production of four recombinant pharmaceutical proteins using different platforms, highlighting from these examples the unique advantages of plant-based systems over traditional fermenter-based expression platforms

Comparative Evaluation of Recombinant Protein Production in Different Biofactories: The Green Perspective

In recent years, the production of recombinant pharmaceutical proteins in heterologous systems has increased significantly. Most applications involve complex proteins and glycoproteins that are difficult to produce, thus promoting the development and improvement of a wide range of production platforms. No individual system is optimal for the production of all recombinant proteins, so the diversity of platforms based on plants offers a significant advantage. Here, we discuss the production of four recombinant pharmaceutical proteins using different platforms, highlighting from these examples the unique advantages of plantbased systems over traditional fermenter-based expression platforms

Comparative Evaluation of Recombinant Protein Production in Different Biofactories: The Green Perspective

In recent years, the production of recombinant pharmaceutical proteins in heterologous systems has increased significantly. Most applications involve complex proteins and glycoproteins that are difficult to produce, thus promoting the development and improvement of a wide range of production platforms. No individual system is optimal for the production of all recombinant proteins, so the diversity of platforms based on plants offers a significant advantage. Here, we discuss the production of four recombinant pharmaceutical proteins using different platforms, highlighting from these examples the unique advantages of plant-based systems over traditional fermenter-based expression platforms

A comparative analysis of recombinant protein expression in different biofactories: bacteria, insect cells and plant systems

Plant-based systems are considered a valuable platform for the production of recombinant proteins as a result of their well-documented potential for the flexible, low-cost production of high-quality, bioactive products. In this study, we compared the expression of a target human recombinant protein in traditional fermenter-based cell cultures (bacterial and insect) with plant-based expression systems, both transient and stable. For each platform, we described the set-up, optimization and length of the production process, the final product quality and the yields and we evaluated provisional production costs, specific for the selected target recombinant protein. Overall, our results indicate that bacteria are unsuitable for the production of the target protein due to its accumulation within insoluble inclusion bodies. On the other hand, plant-based systems are versatile platforms that allow the production of the selected protein at lower-costs than Baculovirus/insect cell system. In particular, stable transgenic lines displayed the highest-yield of the final product and transient expressing plants the fastest process development. However, not all recombinant proteins may benefit from plant-based systems but the best production platform should be determined empirically with a case-by-case approach, as described here

Vanillin accumulation by Vanilla planifolia: what do we think we know and what do we really know?

Natural vanilla is one of the most popular and valuable spice widely employed in food, beverages and cosmetics. This fragrance is produced in fruits of the orchid Vanilla planifolia and, from a chemical point of view, it is composed by a large number of molecules (more than 200), where vanillin, vanillic acid, vanillyl alcohol, p-hydroxybenzaldehyde and p-hydroxybenzoic acid are the most representative. These substances possess important antioxidant capacities and beneficial nutraceutical properties, as demonstrated by several works (Shyamala et al., 2007; Ueno et al., 2019). To overcome the high market demand of vanilla flavor, most of the vanilla commercial products are made of vanillin, obtained by chemical conversion of several substrates, most of which derive from by-products of lignin decomposition processes or from fossil fuels (Mart\u103u et al., 2021). This chemically synthetized vanillin is considered as a non-natural flavor, the resulting aroma covers only partially the wide and complex fragrance typical of the natural spice, and its low commercial value is not comparable to that of natural vanilla. In V. planifolia, vanillin biosynthetic pathway is described to start from ferulic acid and ferulic acid glucoside that would be respectively converted to vanillin and vanillin glucoside by the activity of VANILLIN SYNTHASE (VpVAN). Gallage and coworkers stated the activity of this enzyme through heterologous expression in yeasts and tobacco and suggested that VpVAN, according to its function, is highly expressed in the inner part of the vanilla pods and in particular in the phenyloplasts, compartments deriving from the conversion of chloroplasts during fruit ripening (Gallage et al., 2014; Gallage et al., 2018). However, the results obtained from Gallage and colleagues were questioned in another recent publication, in which the authors failed to produce vanillin by exploiting VpVAN, thus suggesting a re-evaluation of VpVAN role and the existence of more complex and extended pathway(s) for vanillin production in vanilla pods (Yang et al., 2017). In our project, bioinformatics researches and in-vitro experiments suggested that VpVAN is highly expressed in roots, stems, leaves and in-vitro protocorms of V. planifolia, even though these organs and the in vitro culture do not accumulate vanillin. On the other side, feeding the Vp in vitro culture with the putative vanillin precursor ferulic acid did not result in vanillin or vanillin derivatives accumulation, and the same results were obtained with Nicotiana benthamiana and Beta vulgaris transiently expressing the same gene. These evidences support the doubts of Yang and coworkers on the real VpVAN gene function. Thus, we started different approaches as feeding with various putative molecular precursors, transient expression of candidate genes and metabolite detection with high resolution LC-MS