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Purification, crystallization, and preliminary structural analysis of multivalent immunogenic effector protein-anchored SARS-CoV-2 RBD

Kwon, T. H.; Kim, T. G.

2023-08-18 biophysics
10.1101/2023.08.17.553661 bioRxiv
Show abstract

The continuous spread of highly transmissible variants of concern and the potential diminished effectiveness of existing vaccines necessitate ongoing research and development of new vaccines. Immunogenic molecule-anchored antigen has demonstrated superior efficacy in subunit vaccination, primarily due to enhanced cellular uptake facilitated by the affinity between the surface of Immunogenic molecule and the cell membrane. Based on the Immunogenic recombinase B. malayi RecA (BmRecA), we have overexpressed the construct of BmRecA with SARS-CoV-2 RBD (BmRecA-RBD) that exists as a stable helical filament formation; it was purified and crystallized to obtain X-ray diffraction data at 2.7 [A], belonged to the hexagonal symmetry group P65 in the unit-cell parameters of a=b=122.12, c=75.55 and ={beta}=90{degrees}, {gamma}=120{degrees}. The Matthews coefficient was estimated to be 3.12 [A]3 Da-1, corresponding to solvent contents of 52.65.

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