Rapid and Cost-Effective Diagnostic Blot Assays Based on the Use of Plant-Produced Recombinant Antigens: Lessons Learned from the SARS-CoV-2 RBD Antigen
Year: 2025
Type: article
Abstract: 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.
DOI: 10.3390/ijms26104500
Institutions: San Raffaele University of Rome, Istituto di Ricovero e Cura a Carattere Scientifico San Raffaele, Istituti di Ricovero e Cura a Carattere Scientifico, National Agency for New Technologies, Energy and Sustainable Economic Development, ENEA Casaccia Research Centre, CEINGE Biotecnologie Avanzate Franco Salvatore (Italy), Ospedale dell' Angelo, Santissima Trinità, INGV Osservatorio Etneo
Language: English
Subfield: Biotechnology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $3,499
Awards: G1747116307, G2636645585, G7002540278
OpenAlex ID: W4410192199
