We have recently described an efficient transient expression system mediated by Agrobacterium tumefaciens for the production of HIV-1 Nef protein in Nicotiana benthamiana plants. In order to enhance the yield of recombinant protein we assayed the effect of three gene- silencing viral suppressor proteins (P25 of Potato Virus X, P19 of Artichoke Mottled Crinckle virus and Tomato Bushy Stunt virus) on Nef expression levels. Results demonstrated that AMCV-P19 gave the highest Nef yield (1.3% of total soluble protein) and that this effect was correlated to a remarkable decrease of Nef-specific small interfering RNAs (siRNAs) indicating an effective modulation of RNA silencing mechanisms. Here we report additional data on the production of different heterologous proteins including human immunoglobulin heavy and light chains and a virus coat protein that demonstrate the robustness of this co-agroinfiltration expression system boosted by the AMCV-P19 gene-silencing suppressor.

Celiac disease is an immune-mediated disorder caused by the ingestion of gluten proteins. The gluten-free diet is currently the only therapy to achieve the symptoms’ remission. Biotechnological approaches are currently being explored to obtain safer and healthier food for celiacs. This article analyzes consumer awareness and acceptance of advanced biotechnologies to develop gluten-free products. An online snowball sampling questionnaire was proposed to 511 Italian participants, selected among celiac and non-celiac people, from December 2020 to January 2021, during the second wave of the COVID-19 pandemic. Overall, 64% of respondents favor food biotechnology, as long as it has benefits for health or the environment. Moreover, biotechnology perception differs according to education level and type. A total of 65% of the survey participants would taste gluten-free products obtained through a biotechnological approach, and 57% would buy them at a higher price than the current market price. Our results show a change in public opinion about the usefulness of food biotechnology and its moral acceptability compared to 20 years ago. However, the study of public opinion is very complex, dealing with individuals with social, economic, and cultural differences. Undoubtedly, the scientific dissemination of genetic biotechnologies must be more effective and usable to increase the level of citizens’ awareness.

Infectious Bursal Disease Virus (IBDV), the etiological agent of Gumboro disease, causes mortality and immunosuppression in chickens and major losses to poultry industry worldwide. The IBDV major capsid protein VP2 is considered the best candidate for the production of novel subunit vaccines. This structural protein contains the major conformational epitopes responsible for the induction of IBDV neutralizing antibodies in chickens and has been demonstrated able to form supramolecular structures in yeast and insect cells. The aim of this study was to express an engineered version of the VP2 protein (His-pVP2) to verify its ability to self-assemble into virus-like particles in plants. The recombinant VP2 was transiently expressed by agroinfiltration in Nicotiana benthamiana and transmission electron microscopy of sucrose density gradient fractions revealed the presence of a mixed population of differently shaped particles ranging from spherical capsids, with a diameter between ~25 and ~70 nm, to tubular structures, with variable length (from 100 to 400 nm). The recombinant VP2-based particles when used for the intramuscular immunization of specific-pathogen-free chicks resulted able to induce the production of anti-IBDV specific antibodies at titers comparable to those induced by a commercial vaccine. Moreover, all the immunized birds survived to the challenge with a Moroccan very virulent IBDV strain with no major histomorphological alterations of the Bursa of Fabricius, similarly to what obtained with the commercial inactivated vaccine.

The ongoing demand for reliable, cost-effective, and scalable diagnostic solutions during the SARS-CoV-2 pandemic has highlighted the need for innovative production platforms. Here, we present a plant-based molecular farming (PMF) strategy for the production of the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein fused with Fc (RBDw-Fc). The RBDw-Fc antigen was transiently expressed in Nicotiana benthamiana plant, achieving high yields and purity. Antigen-antibody binding assays were used to assess its functionality. The purified antigen was used to develop a rapid diagnostic blot assay to screen both pre and post vaccinated and pre and post infected plasma EDTA samples with high sensitivity and specificity. Our results demonstrate that the RBDw-Fc-based assay is effective for SARS-CoV-2 detection, and offers significant advantages in terms of production speed, scalability, and cost-efficiency compared to traditional systems, such as cell-culture-based production. This kind of diagnostic test deliver accurate results in just few minutes, enhancing its applicability in clinical and resource-limited settings. This study highlights the versatility of PMF as a platform for producing high-quality reagents, with potential applications beyond SARS-CoV-2 diagnostics. The RBDw-Fc antigen-based method offers a model for the development of rapid, economical, and adaptable screening tools for emerging infectious diseases and future pandemics.

The ongoing demand for reliable, cost-effective, and scalable diagnostic solutions during the COVID-19 pandemic emphasized the need for innovative production platforms. In this study, we present a plant-based molecular farming (PMF) strategy for the production of the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein fused with an Fc region (RBDw-Fc). The RBDw-Fc antigen was transiently expressed in the Nicotiana benthamiana plant, achieving high yields and purity. Its functionality was assessed through antigen–antibody binding assays. The purified antigen was subsequently employed in the development of a rapid diagnostic blot assay capable of screening plasma EDTA samples from pre- and post-vaccinated as well as pre- and post-infected individuals, demonstrating high sensitivity and specificity. Our results show that the RBDw-Fc-based assay is effective for SARS-CoV-2 detection and offers considerable advantages in terms of production speed, scalability, and cost efficiency compared to traditional systems, such as cell-culture-based production. The assay delivers accurate results in just a few minutes, making it particularly suitable for clinical and resource-limited settings. This study highlights the versatility of PMF as a platform for producing high-quality reagents, with promising applications beyond SARS-CoV-2 diagnostics. The RBDw-Fc antigen-based method provides a model for the rapid, economical, and flexible development of screening tools for emerging infectious diseases and future pandemics.