IIBIO   27936
Unidad Ejecutora - UE
congresos y reuniones científicas
Horizontal and Collaborative against the clock work in pandemic: Production, characterization and distribution of a low-cost SARS-CoV-2 antigen by the Argentinian AntiCovid Consortium.
Congreso; Reunion Anual de Sociedad Argentina de Biofisica; 2020
Horizontal and Collaborative against the clock work in pandemic: Production, characterization and distribution of a low-cost SARS-CoV-2 antigen by the Argentinian AntiCovid Consortium.Argentinian AntiCovid Consortium: formed by 34 researchers of CONICET working at iB3/AGBT-FBMC-FCEN-UBA; QB-FCEN-UBA; FFyB-UBA; FMed-UBA; UTN; IIB-UNSAM; ICT- Milstein; (anticovid.arg@gmail.com).December 19th 2019, an epidemiological alert came from China to the whole world: A new virus causing Severe Acute Respiratory Syndrome may be evolving. On January 10th the world got to know the genome of the SARS CoV2. Shortly after the installed quarantine period in Argentina, a group of researchers from different institutions and with diverse relevant expertise began a collaborative work to contribute in the development of local accessible high-quality molecular tools against SARS-CoV-2. Our work was mainly aimed at producing the Receptor Binding Domain (RBD) of the SARS-CoV-2 Spike protein at low cost. In this work, we produced RBD from SARS-CoV-2 Spike protein in both the well-established HEK-293T mammalian cells system and also in the yeast Pichia pastoris. P. pastoris is a cost-effective and easily scalable system for recombinant protein production. Importantly, we found that that (i) RBD produced in both systems was properly folded as judged by CD spectroscopy; (ii) mass spectrometry analysis and glycosidase digestion suggested that both forms are differentially glycosylated; (iii) both variants are conformationally stable and their thermal stabilities depend on the ionic strength in the same way, and (iv) antibodies generated in mice immunized with RBD from yeast recognize the protein produced in mammalian cells and vice versa. Simultaneously, the production of RBD in P. pastoris was scaled-up in a stirred-tank bioreactor, with yields above 100 mg/L of recombinant protein (∼90% of purity), thus allowing many collaborations with different projects regarding: the development of an anti SARS-CoV-2 antibody detection kit, hens immunizations and IgY production, vaccine investigations, and monoclonal antibody developments. Also, some insights in the design of an improved antigen with better immunogenic characteristics, compared with those of wild type monomeric RBD, obtained through the covalent coupling of RBD to other proteins will be presented. Acknowledgments: INTA, Bioinnovo SA, UNPaz, UNLP, UNQ, UNCUYO, ANLAP, and UNSAM for collaboration; Oxigeno Central, LANAIS-PRO-EM, Technisys S.A., Masonería Argentina, Asoc. Civil Malmann, UBA and ANPCyT and CONICET for resources and Secr. de investigación, Of. vinculación tecnológica, and FundaCEN, FCEN, UBA. Argentinian AntiCovid Consortium (in alphabetical order with equal contribution of all authors): Claudia R. Arbeitman, Analía Amante, Gabriela Auge, Matías Blaustein, Luis Bredeston, Enrique S. Corapi, Patricio O. Craig, Leandro A. Cossio, Liliana Dain, Cecilia D?Alessio, Fernanda Elias, Natalia B. Fernández, Yamila Gándola, Javier Gasulla, Natalia Gorojovsky, Gustavo E. Gudesblat, María G. Herrera, Lorena I. Ibañez, Tommy Idrovo Hidalgo, Matías Iglesias Randon, Laura Kamenetzky, Alejandro D. Nadra, Diego G. Noseda, Carlos H. Paván, María F. Pavan, María F. Pignataro, Javier Ramirez, Ernesto Roman, Lucas A. M. Ruberto, Natalia Rubinstein, Javier Santos, Francisco Velazquez Duarte, Diana E. Wetzler and Alicia M. Zelada