Journal: Molecules

Article Title: Non-Glycosylated SARS-CoV-2 Omicron BA.5 Receptor Binding Domain (RBD) with a Native-like Conformation Induces a Robust Immune Response with Potent Neutralization in a Mouse Model

doi: 10.3390/molecules29112676

Figure Lengend Snippet: SARS-CoV-2 Omicron BA.5 RBD expression and purification in E. coli . ( a ) Schematics of the sequence location of RBD in the SARS-CoV-2 spike protein. ( b ) Ribbon model of SARS-CoV-2 RBD with disulfide bond pairing. ( c ) SARS-CoV-2 Omicron BA.5 RBD expression and purification protocol in E. coli . ( d ) Binding of the SARS-CoV-2 Omicron BA.5 RBD to the hACE2 using an Octet-N1 Bio-Layer Interferometer. RBD was immobilized on a Ni-NTA sensor chip, and hACE2 was in the mobile phase.

Article Snippet: SARS-CoV-2 RBD (5 μg/mL) was immobilized on the Ni-NTA biosensor for 180 s, followed by an association phase with recombinant hACE2 (purity > 90% SDS-PAGE) (Bioworld Tech, St. Louis Park, MN, USA) diluted in kinetics buffer for 300 s. Dissociation was then carried out in the kinetics buffer for 300 s. Binding affinities were calculated using a 1:1 Langmuir binding model.

Techniques: Expressing, Purification, Sequencing, Binding Assay

Journal: Molecules

Article Title: Non-Glycosylated SARS-CoV-2 Omicron BA.5 Receptor Binding Domain (RBD) with a Native-like Conformation Induces a Robust Immune Response with Potent Neutralization in a Mouse Model

doi: 10.3390/molecules29112676

Figure Lengend Snippet: hACE2 inhibition assay. ( a ) Steps of the hACE2 binding inhibition assay using bio-layer interferometry (BLI). RBD was immobilized on the biosensor chip, followed by antisera binding. hACE2 was loaded for the association and dissociation steps assessed in the kinetic buffer. ( b ) Enlarged figure of hACE2 association and dissociation step. ( c ) Inhibition of RBD binding to hACE2 by E. coli -expressed RBD-immunized antisera of one mouse from each group; “M” indicates the identity of the mouse.

Article Snippet: SARS-CoV-2 RBD (5 μg/mL) was immobilized on the Ni-NTA biosensor for 180 s, followed by an association phase with recombinant hACE2 (purity > 90% SDS-PAGE) (Bioworld Tech, St. Louis Park, MN, USA) diluted in kinetics buffer for 300 s. Dissociation was then carried out in the kinetics buffer for 300 s. Binding affinities were calculated using a 1:1 Langmuir binding model.

Techniques: Inhibition, Binding Assay