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Quantifying Absolute Neutralization Titers against SARS-CoV-2 by a Standardized Virus Neutralization Assay Allows for Cross-Cohort Comparisons of COVID-19 Sera

Kasopefoluwa Y. Oguntuyo, Christian S. Stevens, Chuan-Tien Hung, Satoshi Ikegame, Joshua A. Acklin, Shreyas Kowdle, Jillian C. Carmichael, Hsin-Ping Chiu, Kristopher D. Azarm, Griffin D. Haas, Fatima Amanat, Jéromine Klingler, Ian Baine, Suzanne Arinsburg, Juan C. Bandrés, Mohammed Nure Alam Siddiquey, Robert M. Schilke, Matthew D. Woolard, Hongbo Zhang, J. Andrew Duty, Thomas Kraus, Thomas M. Moran, Domenico Tortorella, Jean K. Lim, Andrea V. Gamarnik, Catarina E. Hioe, Susan Zolla‐Pazner, Stanimir S. Ivanov, Jeremy P. Kamil, Florian Krammer, Benhur Lee

2021mBio83 citationsDOIOpen Access PDF

Abstract

Vaccines and antibody-based therapeutics like convalescent-phase plasma therapy are premised upon inducing or transferring neutralizing antibodies that inhibit SARS-CoV-2 entry into cells. Virus neutralization assays (VNAs) for measuring neutralizing antibody titers (NATs) are an essential part of determining vaccine or therapeutic efficacy. However, such efficacy testing is limited by the inherent dangers of working with the live virus, which requires specialized high-level biocontainment facilities. We therefore developed a standardized replication-defective pseudotyped particle system that mimics the entry of live SARS-CoV-2. This tool allows for the safe and efficient measurement of NATs, determination of other forms of entry inhibition, and thorough investigation of virus entry mechanisms. Four independent labs across the globe validated our standardized VNA using diverse cohorts. We argue that a standardized and scalable assay is necessary for meaningful comparisons of the myriad of vaccines and antibody-based therapeutics becoming available. Our data provide generalizable metrics for assessing their efficacy.

Topics & Concepts

NeutralizationVirologyTiterPotencyNeutralizing antibodyVesicular stomatitis virusAntibodyVirusGlycoproteinPandemicAntibody titerMedicineBiologyImmunologyCoronavirus disease 2019 (COVID-19)DiseaseIn vitroInfectious disease (medical specialty)PathologyBiochemistryMolecular biologySARS-CoV-2 and COVID-19 ResearchCOVID-19 Clinical Research StudiesVirus-based gene therapy research
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