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Evaluation of Cell-Based and Surrogate SARS-CoV-2 Neutralization Assays #MMPMID34288726
Sholukh AM; Fiore-Gartland A; Ford ES; Miner MD; Hou YJ; Tse LV; Kaiser H; Zhu H; Lu J; Madarampalli B; Park A; Lempp FA; St Germain R; Bossard EL; Kee JJ; Diem K; Stuart AB; Rupert PB; Brock C; Buerger M; Doll MK; Randhawa AK; Stamatatos L; Strong RK; McLaughlin C; Huang ML; Jerome KR; Baric RS; Montefiori D; Corey L
J Clin Microbiol 2021[Sep]; 59 (10): e0052721 PMID34288726show ga
Determinants of protective immunity against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection require the development of well-standardized, reproducible antibody assays. This need has led to the emergence of a variety of neutralization assays. Head-to-head evaluation of different SARS-CoV-2 neutralization platforms could facilitate comparisons across studies and laboratories. Five neutralization assays were compared using 40 plasma samples from convalescent individuals with mild to moderate coronavirus disease 2019 (COVID-19): four cell-based systems using either live recombinant SARS-CoV-2 or pseudotyped viral particles created with lentivirus (LV) or vesicular stomatitis virus (VSV) packaging and one surrogate enzyme-linked immunosorbent assay (ELISA)-based test that measures inhibition of the spike protein receptor binding domain (RBD) binding its receptor human angiotensin converting enzyme 2 (hACE2). Vero cells, Vero E6 cells, HEK293T cells expressing hACE2, and TZM-bl cells expressing hACE2 and transmembrane serine protease 2 were tested. All cell-based assays showed 50% neutralizing dilution (ND(50)) geometric mean titers (GMTs) that were highly correlated (Pearson r = 0.81 to 0.89) and ranged within 3.4-fold. The live virus assay and LV pseudovirus assays with HEK293T/hACE2 cells showed very similar mean titers, 141 and 178, respectively. ND(50) titers positively correlated with plasma IgG targeting SARS-CoV-2 spike protein and RBD (r = 0.63 to 0.89), but moderately correlated with nucleoprotein IgG (r = 0.46 to 0.73). ND(80) GMTs mirrored ND(50) data and showed similar correlation between assays and with IgG concentrations. The VSV pseudovirus assay and LV pseudovirus assay with HEK293T/hACE2 cells in low- and high-throughput versions were calibrated against the WHO SARS-CoV-2 IgG standard. High concordance between the outcomes of cell-based assays with live and pseudotyped virions enables valid cross-study comparison using these platforms.
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J+Clin+Microbiol 2021 ; 59 (10): e0052721
