Recombinant Production and Functional Characterization of Andes Virus Gn and Gc in Pichia pastoris: A Path Towards Subunit Vaccine Development

• DOI: 10.20944/preprints202410.0828.v1
• Publication Date: 2024-10-12T00:31:50Z
• Authors: Maria Francisca S...; Claudia Paola Cam...; Belen B.A. Jerez ...; Fátima Reyes; Frank Camacho-Cas...; Jannel Acosta Alba; Raquel Montesino ...; Rafael Maura; Fernando J. Ramos...; Jorge R. Toledo; Oliberto Sánchez ...

Hantavirus Pulmonary Syndrome (HPS), caused by the Andes virus (ANDV), is a significant public health concern in South America, with no available vaccines or specific treatments. Neutralizing antibodies targeting ANDV glycoproteins Gn and Gc have shown protective effects animal models, making them promising vaccine candidates. This study reports recombinant expression, purification, immunogenicity evaluation of sGn sGc produced methylotrophic yeast Pichia pastoris. Initially, was expressed as secretable protein, but it formed aggregates once accumulated culture medium. To address this, same strain transformed vector expressing Gc, based on hypothesis that folding depends presence. However, co-expression resulted both proteins becoming trapped insoluble inside yeast, resembling inclusion bodies seen E. coli. The were solubilized purified using metal ion affinity chromatography, followed refolding step before being used for immunization. antigens, formulated aluminum hydroxide (AlOH), tested Syrian hamsters. immunization results showed sGn-sGc formulation induced strong Th1-biased immune response, characterized high levels IFN-γ, IL-12, IL-6 cytokines, along antibody response against titers exceeding 1:10,000. These findings suggest potential these antigens candidates subunit HPS.