A custom multiplex bead array was designed (Supplemental Table 3) and coupled with SARS-CoV-1 and SARS-CoV-2 spike-1 (stem, Sino Biological), spike-2 (head, ACRO Biosystems), RBD (BEI Resources), and N (ACRO Biosystems), as described previously (45 (link)). In addition, SARS-CoV-2 spike trimers (provided in-house) and SARS-CoV-2 spike trimers (BPS Bioscience) were included. Tetanus toxoid (Sigma-Aldrich), influenza hemagglutinin (H1Cal2009, Sino Biological), and SIV-gp120 (Sino Biological) were included as positive and negative control antigens, respectively. Antigens were covalently coupled to magnetic carboxylated beads (Bio-Rad) using a 2-step carbodiimide reaction and blocked with 0.1% BSA, before being resuspended and stored in PBS with 0.05% sodium azide until use.
Habel J.R., Chua B.Y., Kedzierski L., Selva K.J., Damelang T., Haycroft E.R., Nguyen T.H., Koay H.F., Nicholson S., McQuilten H.A., Jia X., Allen L.F., Hensen L., Zhang W., van de Sandt C.E., Neil J.A., Pragastis K., Lau J.S., Jumarang J., Allen E.K., Amanant F., Krammer F., Wragg K.M., Juno J.A., Wheatley A.K., Tan H.X., Pell G., Walker S., Audsley J., Reynaldi A., Thevarajan I., Denholm J.T., Subbarao K., Davenport M.P., Hogarth P.M., Godfrey D.I., Cheng A.C., Tong S.Y., Bond K., Williamson D.A., McMahon J.H., Thomas P.G., Pannaraj P.S., James F., Holmes N.E., Smibert O.C., Trubiano J.A., Gordon C.L., Chung A.W., Whitehead C.L., Kent S.J., Lappas M., Rowntree L.C, & Kedzierska K. (2023). Immune profiling of SARS-CoV-2 infection during pregnancy reveals NK cell and γδ T cell perturbations. JCI Insight, 8(7), e167157.
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