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The protein-bound resin was extensively washed with 100 mL of buffer consisting of 20 mM Tris-Cl (pH 8.0), 500 mM NaCl, 1 mM DNQX, and 0.02% DDM at room temperature, and then slowly washed with 200 mL of buffer consisting of 20 mM Tris-Cl (pH 8.0), 300 mM NaCl, and 0.02% DDM at room temperature at a rate of approximately 0.3 ml/min

S expression plasmids for current SARS-CoV-2 VoC were similarly codon optimized, modified and included the following mutations compared to WA-1: Beta/B.1.351 (D80A, D215G, del 241-243, K417N, E484K, N501Y, D614G, A701V,), Delta/B.1.617.2 (T19R, E156G, del 157-158, L452R, T478K, D614G, P681R, D950N), Omicron BA.1 (A67V, 69/70, T95I, G142D, del 143-145, N211I, del 212R214, insG339D S371L, S373P, S375F, K417N, N440K, G446S, S477N, T478K, E484A, Q493R, G496S, Q498R, N501Y, Y505H, T547K, D614G, H655Y, N679K, P681H, N764K, D796Y, N856K, Q954H, N969K, L981F), Omicron BA.2 (T19I, L24-, P25-, P26-, A27S, G142D, V213G, G339D, S371F, S373P, S375F, T376A, D405N, R408S, K417N, N440K, S477N, T478K, E484A, Q493R, Q498R, N501Y, Y505H, D614G, H655Y, N679K, P681H, N764K, D796Y, Q954H, N969K), and Omicron BA.5 (T19I, del L24, del P25, del P26, A27S, del H69, del V70, G142D, V213G, G339D, S371F, S373P, S375F, T376A, D405N, R408S, K417N, N440K, L452R, S477N, T478K, E484A, F486V, Q498R, N501Y, Y505H, D614G, H655Y, N679K, P681H, N764K, D796Y, Q954H, N969K) from Genscript

There are three phenolic hydroxyl groups distributed on the A and B rings, while on the C ring there are double bonds and methyl groups