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In this study, we designed and evaluated novel mRNA\u2010based vaccines encoding conserved domains of SARS\u2010CoV\u20102, including the spike S2 subunit, NSP3 Macrodomain I (Mac I), and papain\u2010like protease (PLpro). These mRNAs were synthesized, cloned, and encapsulated into lipid nanoparticles (LNPs) with 70%\u201380% efficiency and favorable physicochemical properties. In vitro characterization showed efficient antigen expression with 60%\u201380% cell survival. Immunization of BALB\/c mice induced strong humoral responses, particularly against S2 and Mac I, marked by IgG titers, class switching, and durable antibody presence up to 84 days post\u2010booster. PLpro elicited minimal antibody responses, likely due to limited stability or intrinsic low immunogenicity. Combinatorial LNPs encoding multiple antigens maintained robust expression and elicited comparable immune responses despite dose splitting. Anti\u2010S2 antibodies effectively inhibited spike protein uptake in vitro. Importantly, the selected antigenic regions exhibited high sequence conservation across major SARS\u2010CoV\u20102 variants, underscoring their broad\u2010spectrum vaccine potential. These findings support further development of domain\u2010targeted, multi\u2010antigen mRNA vaccines to enhance cross\u2010variant protection and T\u2010cell\u2010mediated immunity.<\/jats:p>","DOI":"10.1002\/biot.70294","type":"journal-article","created":{"date-parts":[[2026,8,17]],"date-time":"2026-08-17T06:19:20Z","timestamp":1786947560000},"source":"Crossref","is-referenced-by-count":0,"title":["Immunological Evaluation of mRNA\u2010LNP Vaccines Targeting Conserved S2 and NSP3 Domains of SARS\u2010CoV\u20102"],"prefix":"10.1002","volume":"21","author":[{"given":"Feroza","family":"Begum","sequence":"first","affiliation":[{"name":"CSIR\u2010Indian Institute of Chemical Biology, Jadavpur  Kolkata West Bengal India"},{"name":"Academy of Scientific and Innovative Research (AcSIR)  Ghaziabad India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Md Hasan","family":"Mallik","sequence":"additional","affiliation":[{"name":"CSIR\u2010Indian Institute of Chemical Biology, 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