Design and functional analysis of the glycosylated SARS-CoV-2 RBD.
<p>(A) Design and construction of three mRNA expressing trimerization RBD with glycosylation. SP: signal peptide; T4f motif: T4 fibritin motif; (B) Circular dichroism of three purified RBD proteins; (C, D) Purified RBD proteins detected by SDS-PAGE and Western blot. (E) Western blot analysis o...
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2024
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| Summary: | <p>(A) Design and construction of three mRNA expressing trimerization RBD with glycosylation. SP: signal peptide; T4f motif: T4 fibritin motif; (B) Circular dichroism of three purified RBD proteins; (C, D) Purified RBD proteins detected by SDS-PAGE and Western blot. (E) Western blot analysis of deglycosylated RBD proteins; (F) Purified RBD proteins detected by reducing and non-reducing page. The left line of each protein was subjected to 4–12% protein gel electrophoresis after boiling in 5×SDS-PAGE loading buffer, which disrupts the trimeric configuration, and the right line of each protein was subjected to 4–12% protein gel electrophoresis after mixed with 5×native sample loading buffer, preserving the protein’s oligomeric structure; (G) The affinity analysis of the monoclonal antibody B38 with different variants of RBD proteins and deglycosylated RBD. WT: wide-type, M1: Mutation 1, M2: Mutation 2.</p> |
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