Journal: medRxiv

Article Title: Development of an Optical Assay to Detect SARS-CoV-2 Spike Protein Binding Interactions with ACE2 and Disruption of these Interactions Using Electric Current

doi: 10.1101/2020.11.24.20237628

Figure Lengend Snippet: Schematic illustration of SARS-CoV-2 spike protein and ACE2 receptor binding. (a) SARS-CoV-2 binding to the ACE2 receptor on the host cell surface. (b) Binding of ACE2 and spike protein along with illustration of the spike protein subunits, S1 and S2. (c) Schematic showing the distribution of the spike protein in solution. (d) Schematic showing the distribution of ACE2 in suspension. (e) Distribution of ACE2 and S protein after binding in solution.

Article Snippet: Two different spike S1 recombinant proteins (S1X (ProSci Cat no. 10-109) and S1Y (ProSci Cat no. 10-111)) were also purchased in the lyophilized form.

Techniques: Binding Assay

Journal: medRxiv

Article Title: Development of an Optical Assay to Detect SARS-CoV-2 Spike Protein Binding Interactions with ACE2 and Disruption of these Interactions Using Electric Current

doi: 10.1101/2020.11.24.20237628

Figure Lengend Snippet: Optical measurements of the spike protein subunits S1 and S2: (a) Measured responses for spikes proteins S1 and S2 at the highest concentration individually (S1B and S2B, respectively), along with their corresponding blanks. (b) Time domain measurements of the microcentrifuge tube, the blank (shown in gray circles) versus water (red circles) at a wavelength of 623 nm. (c) Measured optical responses for the mixed protein samples versus time. Samples S1B and S2B were at 5000 copies per ml, S2C, S2D, S2E and S2F are the serial dilutions of S2B at 10-, 100-, 1,000- and 10,000-fold, respectively. (d) Relative change in light intensity per light path versus loaded mass. All optical responses were measured at 623 nm. Light intensity was measured as arbitrary units (a.u).

Article Snippet: Two different spike S1 recombinant proteins (S1X (ProSci Cat no. 10-109) and S1Y (ProSci Cat no. 10-111)) were also purchased in the lyophilized form.

Techniques: Concentration Assay

Journal: medRxiv

Article Title: Development of an Optical Assay to Detect SARS-CoV-2 Spike Protein Binding Interactions with ACE2 and Disruption of these Interactions Using Electric Current

doi: 10.1101/2020.11.24.20237628

Figure Lengend Snippet: Optical detection of binding interactions between ACE2 and other proteins. (a) Measured light intensities over time for individual assessment of ACE2, S1X, S1Y, and BSA. (b) The measured mixed light intensities versus time for ACE2 mixed with either S1X, or S1Y, or BSA. (c) The measured ACE2-S1X interaction profile for an extended time period.

Article Snippet: Two different spike S1 recombinant proteins (S1X (ProSci Cat no. 10-109) and S1Y (ProSci Cat no. 10-111)) were also purchased in the lyophilized form.

Techniques: Binding Assay

Journal: medRxiv

Article Title: Development of an Optical Assay to Detect SARS-CoV-2 Spike Protein Binding Interactions with ACE2 and Disruption of these Interactions Using Electric Current

doi: 10.1101/2020.11.24.20237628

Figure Lengend Snippet: Schematic illustration of SARS-CoV-2 spike protein and ACE2 receptor binding. (a) SARS-CoV-2 binding to the ACE2 receptor on the host cell surface. (b) Binding of ACE2 and spike protein along with illustration of the spike protein subunits, S1 and S2. (c) Schematic showing the distribution of the spike protein in solution. (d) Schematic showing the distribution of ACE2 in suspension. (e) Distribution of ACE2 and S protein after binding in solution.

Article Snippet: The SARS-CoV-2 S1 (ProSci, Cat.No.97-087) and S2 (ProSci Cat.No.97-079) recombinant proteins were purchased in lyophilized form and resuspended according to the manufacturer’s instructions at a stock concentration of 600 µg/ml for S1 and 200 µg/ml for S2 protein in sterile water.

Techniques: Binding Assay

Journal: medRxiv

Article Title: Development of an Optical Assay to Detect SARS-CoV-2 Spike Protein Binding Interactions with ACE2 and Disruption of these Interactions Using Electric Current

doi: 10.1101/2020.11.24.20237628

Figure Lengend Snippet: Optical detection of binding interactions between ACE2 and other proteins. (a) Measured light intensities over time for individual assessment of ACE2, S1X, S1Y, and BSA. (b) The measured mixed light intensities versus time for ACE2 mixed with either S1X, or S1Y, or BSA. (c) The measured ACE2-S1X interaction profile for an extended time period.

Article Snippet: Two different spike S1 recombinant proteins (S1X (ProSci Cat no. 10-109) and S1Y (ProSci Cat no. 10-111)) were also purchased in the lyophilized form.

Techniques: Binding Assay