Production and characterization of the SARS-CoV-2 receptor-binding domain expressed in Lactuca sativa

The emergence of coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), presents a critical challenge to global public health. Despite the pressing need for scalable and affordable vaccines, traditional vaccine production is impeded by high costs, reliance on cold-chain logistics, and limited accessibility in resource-poor regions. Plant-based expression systems, such as Lactuca sativa (lettuce), offer a promising alternative for producing recombinant antigens, including the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein. This study optimises the production and functional characterisation of the SARS-CoV-2 RBD expressed in Lactuca sativa to assess its potential as a plant-derived vaccine. The RBD antigen was selected, and its expression cassette was designed and constructed using SnapGene software. The RBD was cloned into a pRI101-AN vector and transiently expressed in Lactuca sativa via agroinfiltration. Expression was confirmed by PCR and immunoassays (ELISA). Immunogenicity testing was conducted by orally administering harvested Lactuca sativa leaves to specialised pathogen-free (SPF) female BALB/c mice via gastric intubation. Transient expression in Lactuca sativa yielded approximately 1.5 mg of RBD protein per gram of fresh leaf weight. Immunized mice produced SARS-CoV-2-specific IgA and IgG antibodies targeting the RBD region, demonstrating successful immunogenicity. These findings highlight the potential of Lactuca sativa as a platform for developing plant-based vaccines against SARS-CoV-2. This approach could significantly contribute to combating the escalating endemicity of COVID-19 and align with Malaysia’s efforts to produce innovative vaccines locally. However, further evaluations are necessary to validate its safety and efficacy for broader applications.

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