ace2  (R&D Systems)

Journal: Process Biochemistry (Barking, London, England)

Article Title: Process development of a SARS-CoV-2 nanoparticle vaccine

doi: 10.1016/j.procbio.2023.03.014

Figure Lengend Snippet: Principle of the potency binding assay. When Spike RBD-Fc and ACE2-biotin bind to each other, the streptavidin donor beads, and the protein A acceptor beads are brought into close proximity. When a laser is applied to this complex, ambient oxygen is converted to singlet oxygen by the donor beads. If the beads are in close proximity, an energy transfer from the singlet oxygen to the thioxene derivative in the acceptor beads occurs, resulting in the production of luminescence. Component A binds to ACE2-biotin and prevents it from binding to Spike RBD-Fc, thereby decreasing the luminescence output in a dose-dependent manner.

Article Snippet: Component A dilutions (20 μL) over the range of 3000 to 0.786 ng/mL were incubated with 20 μL of recombinant human biotinylated ACE2 (ACE2-biotin) (Acro BioSystems) at 80 ng/mL, 20 μL of recombinant SARS-CoV-2 (COVID-19) S protein RBD, Fc Tag (Spike RBD-Fc) (Acro BioSystems) at 40 ng/mL, and 40 μL of the AlphaScreen® General IgG (Protein A) Detection Kit containing Streptavidin donor beads and Protein A acceptor beads (PerkinElmer) at 0.01 mg/mL in half area 96-well plates (Corning).

Techniques: Binding Assay

Journal: Process Biochemistry (Barking, London, England)

Article Title: Process development of a SARS-CoV-2 nanoparticle vaccine

doi: 10.1016/j.procbio.2023.03.014

Figure Lengend Snippet: Representative dose response curve in the potency binding assay. Increasing concentrations of Component A inhibit binding of ACE2-biotin to Spike RBD-Fc resulting in a dose-dependent decrease in luminescence.

Article Snippet: Component A dilutions (20 μL) over the range of 3000 to 0.786 ng/mL were incubated with 20 μL of recombinant human biotinylated ACE2 (ACE2-biotin) (Acro BioSystems) at 80 ng/mL, 20 μL of recombinant SARS-CoV-2 (COVID-19) S protein RBD, Fc Tag (Spike RBD-Fc) (Acro BioSystems) at 40 ng/mL, and 40 μL of the AlphaScreen® General IgG (Protein A) Detection Kit containing Streptavidin donor beads and Protein A acceptor beads (PerkinElmer) at 0.01 mg/mL in half area 96-well plates (Corning).

Techniques: Binding Assay