A COVID-19 vaccine candidate based on SARS-CoV-2 spike protein and immune-stimulating complexes

VILLARRAZA, JAVIER; FUSELLI, ANTONELA; GUGLIOTTA, AGUSTINA; GARAY, ERNESTO; RODRÍGUEZ, MARÍA CELESTE; FONTANA, DIEGO; ANTUÑA, SEBASTIÁN; GASTALDI, VICTORIA; BATTAGLIOTTI, JUAN MANUEL; TARDIVO, MARÍA BELÉN; ALVAREZ, DIEGO; CASTRO, ELIANA; CASSATARO, JULIANA; CEAGLIO, NATALIA; PRIETO, CLAUDIO

APPLIED MICROBIOLOGY AND BIOTECHNOLOGY

SPRINGER

Año: 2023 vol. 107 p. 3429 - 3441

0175-7598

Abstract: Spike protein from SARS-CoV-2, the etiologic agent of the COVID-19 pandemic disease, constitutes a structural protein that proved to be the main responsible for neutralizing antibody production. Thus, its sequence is highly considered for the design of candidate vaccines. Animal cell culture represents the best option for the production of subunit vaccines based on recombinant proteins since they introduce post-translational modifications that are important to mimic the natural antigenic epitopes. Particularly, the human cell line HEK293T has been explored and used for the production of biotherapeutics since the products derived from them present human-like post-translational modifications that are important for the protein’s activity and immunogenicity. The aim of this study was to produce and characterize a potential vaccine for COVID-19 based on the spike ectodomain (S-ED) of SARS-CoV-2 and two different adjuvants: aluminum hydroxide (AH) and immune-stimulating complexes (ISCOMs). The S-ED was produced in sHEK293T cells using a 1-L stirred tank bioreactor operated in perfusion mode and purified. S-ED characterization revealed the expected size and morphology. High N-glycan content was confirmed. S-ED-specific binding with the hACE2 (human angiotensin-converting enzyme 2) receptor was verified. The immunogenicity of S-ED was evaluated using AH and ISCOMs. Both formulations demonstrated the presence of anti-RBD antibodies in the plasma of immunized mice, being significantly higher for the latter adjuvant. Also, higher levels of IFN-γ and IL-4 were detected after the ex vivo immune stimulation of spleen-derived MNCs from ISCOMs immunized mice. Further analysis confirmed that S-ED/ISCOMs elicit neutralizing antibodies against SARS-CoV-2. Key points: Trimeric SARS-CoV-2 S-ED was produced in stable recombinant sHEK cells in serum-free medium.A novel S-ED vaccine formulation induced potent humoral and cellular immunity.S-ED formulated with ISCOMs adjuvant elicited a highly neutralizing antibody titer.