19n01, a Broadly Neutralizing Antibody Against Omicron BA.1, BA.2, BA.4/5, and Other SARS-CoV-2 Variants of Concern

García-Vega, Melissa; Melgoza-González, Edgar Alonso; Hernández-Valenzuela, Sofía; Hinojosa-Trujillo, Diana; Reséndiz-Sandoval, Mónica; Llamas-Covarrubias, Mara Anaís; Loza-Lopez, Martín; Valenzuela, Olivia; Soto-Gaxiola, Alan; Hernández‐Oñate, Miguel Ángel; Mata-Haro, Verónica; Cassaniti, Irene; Sammartino, José Camilla; Ferrari, Alessandro; Simonelli, Luca; Pedotti, Mattia; Sun, Rui; Zuo, Fanglei; Baldanti, Fausto; Varani, Luca; Marcotte, Harold; Pan‐Hammarström, Qiang; Hernández, Jesús

doi:10.2139/ssrn.4232464

Cited by 1 publication

(1 citation statement)

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“…69728-HRP) were used for one-step detection. The chimeric monoclonal antibody 20n01 specific against SARS-CoV-2 S was kindly donated by the laboratory of Dr. Jesús Hernandez (CIAD, Mexico) 59 and used as a primary antibody, followed by a rabbit anti-human IgG HRP (Abcam, cat. ab6759).…”

Section: Methodsmentioning

confidence: 99%

A pseudovirus-based platform to measure neutralizing antibodies in Mexico using SARS-CoV-2 as proof-of-concept

Cruz-Cardenas

Gutiérrez

López-Arredondo

et al. 2022

Sci Rep

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The gold-standard method to evaluate a functional antiviral immune response is to titer neutralizing antibodies (NAbs) against a viral pathogen. This is historically performed using an in vitro assay of virus-mediated infection, which requires BSL-3 facilities. As these are insufficient in Latin American countries, including Mexico, scant information is obtained locally about viral pathogens NAb, using a functional assay. An alternative solution to using a BSL-3 assay with live virus is to use a BSL-2-safe assay with a non-replicative pseudovirus. Pseudoviral particles can be engineered to display a selected pathogen’s entry protein on their surface, and to deliver a reporter gene into target cells upon transduction. Here we comprehensively describe the first development of a BSL-2 safe NAbs-measuring functional assay in Mexico, based on the production of pseudotyped lentiviral particles. As proof-of-concept, the assay is based on Nanoluc luciferase-mediated luminescence measurements from target cells transduced with SARS-CoV-2 Spike-pseudotyped lentiviral particles. We applied the optimized assay in a BSL-2 facility to measure NAbs in 65 serum samples, which evidenced the assay with 100% sensitivity, 86.6% specificity and 96% accuracy. Overall, this is the first report of a BSL-2 safe pseudovirus-based functional assay developed in Mexico to measure NAbs, and a cornerstone methodology necessary to measure NAbs with a functional assay in limited resources settings.

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Section: Methodsmentioning

confidence: 99%