High-throughput detection of neutralizing antibodies to SARS-CoV-2 variants using flow cytometry
Author:
Zhang Xiaohan, Wang Yajie, Li Mansheng, Li Haolong, Zhang Xiaomei, Xu Xingming, Hu Di, Liang Te, Zhu Yunping, Li Yongzhe, Wang Bingwei, Yu XiaoboORCID
Abstract
AbstractDetecting neutralizing antibodies (NAbs) to SARS-CoV-2 variants is crucial for controlling the spread of COVID-19. In this work, we developed a high-throughput assay for the broad systematic examination of NAbs to eleven SARS-CoV variants of concern (VOCs), which include D614G, Alpha, Beta, Gamma, Delta, Kappa, and Omicron sub-lineages BA.1, BA.2, BA.3, BA.4, and BA.5. The assay is cost-effective, reliable, 35-fold more sensitive than Luminex technology, and can include the new variants during SARS-CoV-2 evolution. Importantly, our results highly correlated with a commercial IgG serological assay (R = 0.89) and cPass, a U.S. FDA-approved surrogate virus neutralization test (sVNT) assay (R = 0.93). With our high-throughput NAb platform, we constructed a comprehensive overview of the interactions between SARS-CoV-2 VOCs’ Spike trimer proteins and ACE2 receptors, leading to the identification of a monoclonal Ab with broad neutralizing activity. Furthermore, when compared to the D614G variant, we found that the serum NAbs elicited by the third dose vaccine (administered after 28 days) demonstrated decreased inhibition to multiple SARS-CoV-2 variants, including Gamma (0.94×), Alpha (0.91×), Delta (0.91×), Beta (0.81×), Kappa (0.81×), BA.2 (0.44×), BA.1 (0.43×), BA.3 (0.41×), BA.5 (0.35×) and BA.4 (0.33×), in cohort of 56 vaccinated individuals. Altogether, our proteomics platform proves to be an effective tool to detect broad NAbs in the population and aid in the development of future COVID-19 vaccines and vaccination strategies.
Publisher
Cold Spring Harbor Laboratory
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