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Quantification of SARS-CoV-2 neutralizing antibody by wild-type plaque reduction neutralization, microneutralization and pseudotyped virus neutralization assays.
Bewley, Kevin R; Coombes, Naomi S; Gagnon, Luc; McInroy, Lorna; Baker, Natalie; Shaik, Imam; St-Jean, Julien R; St-Amant, Natalie; Buttigieg, Karen R; Humphries, Holly E; Godwin, Kerry J; Brunt, Emily; Allen, Lauren; Leung, Stephanie; Brown, Phillip J; Penn, Elizabeth J; Thomas, Kelly; Kulnis, Greg; Hallis, Bassam; Carroll, Miles; Funnell, Simon; Charlton, Sue.
  • Bewley KR; National Infection Service, Public Health England, Salisbury, UK. Kevin.Bewley@phe.gov.uk.
  • Coombes NS; National Infection Service, Public Health England, Salisbury, UK.
  • Gagnon L; Nexelis, Laval, Quebec, Canada.
  • McInroy L; National Infection Service, Public Health England, Salisbury, UK.
  • Baker N; National Infection Service, Public Health England, Salisbury, UK.
  • Shaik I; National Infection Service, Public Health England, Salisbury, UK.
  • St-Jean JR; Nexelis, Laval, Quebec, Canada.
  • St-Amant N; Nexelis, Laval, Quebec, Canada.
  • Buttigieg KR; National Infection Service, Public Health England, Salisbury, UK.
  • Humphries HE; National Infection Service, Public Health England, Salisbury, UK.
  • Godwin KJ; National Infection Service, Public Health England, Salisbury, UK.
  • Brunt E; National Infection Service, Public Health England, Salisbury, UK.
  • Allen L; National Infection Service, Public Health England, Salisbury, UK.
  • Leung S; National Infection Service, Public Health England, Salisbury, UK.
  • Brown PJ; National Infection Service, Public Health England, Salisbury, UK.
  • Penn EJ; National Infection Service, Public Health England, Salisbury, UK.
  • Thomas K; National Infection Service, Public Health England, Salisbury, UK.
  • Kulnis G; Nexelis, Laval, Quebec, Canada.
  • Hallis B; National Infection Service, Public Health England, Salisbury, UK.
  • Carroll M; National Infection Service, Public Health England, Salisbury, UK.
  • Funnell S; National Infection Service, Public Health England, Salisbury, UK.
  • Charlton S; National Infection Service, Public Health England, Salisbury, UK.
Nat Protoc ; 16(6): 3114-3140, 2021 06.
Article in English | MEDLINE | ID: covidwho-1203437
ABSTRACT
Virus neutralization assays measure neutralizing antibodies in serum and plasma, and the plaque reduction neutralization test (PRNT) is considered the gold standard for measuring levels of these antibodies for many viral diseases. We have developed procedures for the standard PRNT, microneutralization assay (MNA) and pseudotyped virus neutralization assay (PNA) for severe acute respiratory syndrome coronavirus 2. The MNA offers advantages over the PRNT by reducing assay time, allowing increased throughput and reducing operator workload while remaining dependent upon the use of wild-type virus. This ensures that all severe acute respiratory syndrome coronavirus 2 antigens are present, but Biosafety Level 3 facilities are required. In addition to the advantages of MNA, PNA can be performed with lower biocontainment (Biosafety Level 2 facilities) and allows for further increases in throughput. For each new vaccine, it is critical to ensure good correlation of the neutralizing activity measured using PNA against the PRNT or MNA. These assays have been used in the development and licensure of the ChAdOx1 nCoV-19 (AstraZeneca; Oxford University) and Ad26.COV2.S (Janssen) coronavirus disease 2019 vaccines and are critical for demonstrating bioequivalence of future vaccines.
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Full text: Available Collection: International databases Database: MEDLINE Document Type: Article Main subject: Neutralization Tests / Antibodies, Neutralizing / SARS-CoV-2 / COVID-19 / Antibodies, Viral Subject: Neutralization Tests / Antibodies, Neutralizing / SARS-CoV-2 / COVID-19 / Antibodies, Viral Language: English Journal: Nat Protoc Clinical aspect: Therapy Year: 2021

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Full text: Available Collection: International databases Database: MEDLINE Document Type: Article Main subject: Neutralization Tests / Antibodies, Neutralizing / SARS-CoV-2 / COVID-19 / Antibodies, Viral Subject: Neutralization Tests / Antibodies, Neutralizing / SARS-CoV-2 / COVID-19 / Antibodies, Viral Language: English Journal: Nat Protoc Clinical aspect: Therapy Year: 2021
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