ppat.1009352.s005.tif (342.93 kB)
The binding activity of anti-SARS-CoV-2 S2 antibodies with spike-expressed MDCK cells in the flow cytometry.
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posted on 2021-02-26, 18:43 authored by Kuan-Ying A. Huang, Tiong Kit Tan, Ting-Hua Chen, Chung-Guei Huang, Ruth Harvey, Saira Hussain, Cheng-Pin Chen, Adam Harding, Javier Gilbert-Jaramillo, Xu Liu, Michael Knight, Lisa Schimanski, Shin-Ru Shih, Yi-Chun Lin, Chien-Yu Cheng, Shu-Hsing Cheng, Yhu-Chering Huang, Tzou-Yien Lin, Jia-Tsrong Jan, Che Ma, William James, Rodney S. Daniels, John W. McCauley, Pramila Rijal, Alain R. TownsendWe produced MDCK-Spike by stably transducing parental MDCK-SIAT1 cells with cDNA expressing full-length SARS-CoV-2 spike glycoprotein. This cell membrane-bound full-length spike carries trimer-stabilizing proline mutations (986KV987 to 986PP987) and substitutions at the S1-S2 furin cleavage site (682RRAR685 to 682GSAG685), which indicates that the spike would display the prefusion conformation. MDCK-H3 cells were stained in the control experiment. Anti-influenza H3 MAb BS-1A was included as an antibody control. Each experiment was repeated twice (n = 2). The binding percentage was presented as mean ± standard error of the mean.
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3 weeksantibody responseSARS-CoV -2 infectionACE 2-blocking responsePotent humoral responsesCOVID -19 patientscross-inhibiting clustersneutralising antibodiesplasmablast-derived IgGAnti-RBD MAbsplasmablast responses32 anti-spike MAbsvirus spike glycoproteinACE 2non-competing pairsplasmablast responsememory B cellsanti-non-RBD S 1plasmablast-derived MAbsanti-receptor-binding domainACE 2-blocking anti-RBD antibodies
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