Working at the interface of biophysics and immunology with strategic applications of the S-RBD from SARS-CoV-2

TOMAS GRAU, RH; PLOPER, D; NAVARRO, SA; ÁVILA, C; STAGNETTO, A; GONZÁLEZ LIZARRAGA, F; SOCÍAS, B; CAZORLA, S; MALDONADO GALEANO, C; PERDIGÓN, G; APICHELA, SILVANA A.; ARGAÑARAZ, MARTÍN E; ZAMPINI, RENATO; VELEZ, E; CHAVES. S; VERA PINGUITORE, E; LLAPUR, C; APFELBAUM, G; AGUILAR LOPEZ. M; ASNAR, P; FLORES, L; TOLEDO, R; COSTA, D; CHEHIN R

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Jornada; Primeras Jornadas Virtuales SAB 2020; 2020

SOCIEDAD ARGENTINA DE BIOFISICA

COVID-19, the pandemic caused by the SARS-CoV-2 virus, threatens to collapse healthcare systems around the world. Therefore, the search for strategies capable of counteracting the effects of this disease is crucial. Taking into account that the receptor binding domain (S-RBD), located in the glycoprotein S of the virus surface is crucial for its pathogenesis, and its region is a critical target for neutralizing antibodies, we set out to express and purify the recombinant S-RBD protein from SARS-CoV-2 and use it for the development of local diagnostic methods and research purposes. To obtain the S-RBD protein, we generated a stable cell line by transducing HEK293T cells using a lentivirus system. The correct transduction and expression were assessed by an Immunofluorescence. S-RBD was purified by FPLC by affinity purification for it subsequent use in: i) the development of an ELISA test ii) immunization of a llama to obtain nanoantibodies and iii) studying the potential effect of SARS-CoV-2 in neurodegenerative diseases. The continuous production of S-RBD protein was used for the development of a reproducible, low cost, sensitive and robust serological test that correlates with neutralizing antibodies. We found that 25% of patients tested with detectable IgG anti-RBD have a high enough antibody titer (above 1/1000) to beconsidered as plasma donors. In llama, the titers obtained were above 1/80000, suitable to continue the project to purify nanoantibodies. The continuous production of S-RBD protein also ensure enough reagent to study how SARS-CoV-2 might affect neurodegeneration from a biochemical and cell biology perspective