Biosynthesis and Immunological Evaluation of a Dual-Antigen Nanoconjugate Vaccine Targeting Group A Streptococcus

<b>Background</b>: Group A Streptococcus (GAS) induces a wide spectrum of human diseases, ranging from superficial infections to life-threatening invasive conditions and post-infectious sequelae such as rheumatic heart disease, posing a heavy global health burden. Critically, there is still no licensed commercial vaccine against GAS, making the development of novel, effective vaccines against this pathogen an urgent and crucial unmet medical need. <b>Methods</b>: We developed a dual-antigen nanoconjugate vaccine against GAS. The Group A Carbohydrate polyrhamnose backbone (GAC^PR) and truncated SLO were site-specifically conjugated via Protein Glycan Coupling Technology (PGCT) in engineered <i>E. coli</i>, and then linked to ferritin nanoparticles using the SnoopTag/SnoopCatcher system. Safety, immunogenicity, and protective efficacy were evaluated in murine models. <b>Results</b>: The nanovaccine was successfully synthesized with high purity. It elicited robust GAC- and SLO-specific IgG/IgG1 responses, conferred 90% survival against lethal GAS challenge (vs. 0-50% in controls), reduced bacterial loads in organs, and lowered inflammatory cytokines. Passive immunization with vaccine-induced serum also achieved 90% survival. No abnormal biochemical indicators, inflammatory responses, or organ pathology were observed. <b>Conclusions</b>: This study successfully developed a bivalent nanoparticle vaccine against GAS. This novel nanovaccine exhibits excellent safety, strong immunogenicity, and effective protection against GAS, providing a promising vaccine candidate.