Quantification of SARS-CoV-2 neutralizing antibody by wild-type plaque reduction neutralization, microneutralization and pseudotyped virus neutralization assays

Kevin R. Bewley, Naomi S. Coombes, Luc Gagnon, Lorna McInroy, Natalie Baker, Imam Shaik, Julien R. St-Jean, Natalie St-Amant, Karen R. Buttigieg, Holly E. Humphries, Kerry J. Godwin, Emily Brunt, Lauren Allen, Stephanie Leung, Phillip J. Brown, Elizabeth J. Penn, Kelly Thomas, Greg Kulnis, Bassam Hallis, Miles Carroll, Simon Funnell, Sue Charlton

Published: 2021-04-22 DOI: 10.1038/s41596-021-00536-y

SARS-CoV-2

neutralizing antibody

PRNT

MNA

PNA

AI 解读

Extended

Extended Data Fig. 1

Qualification findings for microneutralization assay (MNA).

Extended Data Fig. 2

Qualification findings for pseudotyped virus neutralization assay (PNA).

Extended Data Fig. 3 Showing correlation (MiniTab v19) of the ND 50 values of panel of 37 convalescent human sera tested in both PRNT and MNA.

The two assays show excellent correlation (Pearson correlation r= 0.963; p < 0.001) given that both assays are wild-type virus bioassays performed at BSL3.

Extended Data Fig. 4 Showing correlation (MiniTab v19) of the ND 50 values of panel of 37 convalescent human sera tested in both PRNT and PNA.

The two assays show good correlation (Pearson correlation r= 0.862; p < 0.001) given that the PRNT is a wild-type virus assay performed at BSL3 with plaques as readout and the other is performed at BSL2 with a pseudotyped virus with a luminescent readout.

Extended Data Fig. 5 Illustration of poor and strongly neutralizing sera in MNA.

a , Illustration of a poorly neutralizing/negative (green) serum where the neutralization fails to achieve 50% of the virus-only control (VOC) at even the lowest dilution tested; b , Illustration of a strongly neutralizing/highly positive (green) serum where the neutralization fails to cross 50% of the virus-only control (VOC) at the highest dilution tested.

Extended Data Fig. 6

Volumes required for setup of virus dilutions for fine-sighting experiment of wild-type SARS-CoV-2 prior to PRNT assay.

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Quantification of SARS-CoV-2 neutralizing antibody by wild-type plaque reduction neutralization, microneutralization and pseudotyped virus neutralization assays

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