Murine monoclonal antibodies against RBD of the SARS-CoV-2 spike protein as useful analytical tools for subunit vaccine development and clinical trials.

Autor: Blanco OR; Center for Genetic Engineering and Biotechnology of Sancti Spíritus, Circunvalante Norte, Olivos III, Sancti Spíritus, Cuba., Dorta D; Center for Genetic Engineering and Biotechnology of Sancti Spíritus, Circunvalante Norte, Olivos III, Sancti Spíritus, Cuba., Hernández CA; Center for Genetic Engineering and Biotechnology of Sancti Spíritus, Circunvalante Norte, Olivos III, Sancti Spíritus, Cuba., Abreu D; Center for Genetic Engineering and Biotechnology of Sancti Spíritus, Circunvalante Norte, Olivos III, Sancti Spíritus, Cuba., Domínguez AG; Center for Genetic Engineering and Biotechnology of Sancti Spíritus, Circunvalante Norte, Olivos III, Sancti Spíritus, Cuba., Luna Y; Center for Genetic Engineering and Biotechnology of Sancti Spíritus, Circunvalante Norte, Olivos III, Sancti Spíritus, Cuba., Valdivia O; Center for Genetic Engineering and Biotechnology of Sancti Spíritus, Circunvalante Norte, Olivos III, Sancti Spíritus, Cuba., Pérez-Bernal M; Center for Genetic Engineering and Biotechnology of Sancti Spíritus, Circunvalante Norte, Olivos III, Sancti Spíritus, Cuba., Tamayo C; Center for Genetic Engineering and Biotechnology of Sancti Spíritus, Circunvalante Norte, Olivos III, Sancti Spíritus, Cuba., Lemos G; Center for Genetic Engineering and Biotechnology, Ave. 31 e/ 158 y 190, Playa, Havana, Cuba., Pasarón IM; Center for Genetic Engineering and Biotechnology of Sancti Spíritus, Circunvalante Norte, Olivos III, Sancti Spíritus, Cuba., Pérez JJ; Center for Genetic Engineering and Biotechnology of Sancti Spíritus, Circunvalante Norte, Olivos III, Sancti Spíritus, Cuba., Benítez L; Center for Genetic Engineering and Biotechnology of Sancti Spíritus, Circunvalante Norte, Olivos III, Sancti Spíritus, Cuba., Bequet-Romero M; Center for Genetic Engineering and Biotechnology, Ave. 31 e/ 158 y 190, Playa, Havana, Cuba., Fragas A; Civilian Defense Scientific Research Center, Carretera de Jamaica y Autopista 15 Nacional, San José de las Lajas, Mayabeque, Cuba., Cabrera Y; Center for Genetic Engineering and Biotechnology of Sancti Spíritus, Circunvalante Norte, Olivos III, Sancti Spíritus, Cuba. Electronic address: yeosvany.cabrera@cigb.edu.cu., Pérez ER; Center for Genetic Engineering and Biotechnology of Sancti Spíritus, Circunvalante Norte, Olivos III, Sancti Spíritus, Cuba.
Jazyk: angličtina
Zdroj: Journal of immunological methods [J Immunol Methods] 2022 Jan; Vol. 500, pp. 113195. Date of Electronic Publication: 2021 Nov 26.
DOI: 10.1016/j.jim.2021.113195
Abstrakt: COVID-19 pandemic poses a serious threat to human health; it has completely disrupted global stability, making vaccine development an important goal to achieve. Monoclonal antibodies play an important role in subunit vaccines strategies. In this work, nine murine MAbs against the RBD of the SARS-CoV-2 spike protein were obtained by hybridoma technology. Characterization of purified antibodies demonstrated that five of them have affinities in the order of 10 8  L/mol. Six MAbs showed specific recognition of different recombinant RBD-S antigens in solution. Studies of the additivity index of anti-RBD antibodies, by using a novel procedure to determine the additivity cut point, showed recognition of at least five different epitopes. The MAbs CBSSRBD-S.11 and CBSSRBD-S.8 revealed significant neutralizing capacity against SARS-CoV-2 in an ACE2-RBD binding inhibition assay (IC 50  = 85.5pM and IC 50  = 122.7pM, respectively) and in a virus neutralizing test with intact SARS-CoV-2 (VN 50  = 0.552 nM and VN 50  = 4.854 nM, respectively) when D614G strain was used to infect Vero cells. Also CBSSRBD-S.11 neutralized the SARS-CoV-2 strains Alpha and Beta: VN 50  = 0.707 nM and VN 50  = 0.132 nM, respectively. The high affinity CBSSRBD-S.8 and CBSSRBD-S.7 recognized different epitopes, so they are suitable for the development of a sandwich ELISA to quantitate RBD-S recombinant antigens in biomanufacturing processes, as well as in pharmacokinetic studies in clinical and preclinical trials.
(Copyright © 2021. Published by Elsevier B.V.)
Databáze: MEDLINE
Externí odkaz: