CoviDrop SARS-CoV-2 Specific Furin Cleavage Site (P681R Mutation) Blocker/Inhibitor Screening Kit

Product Description

The CoviDrop™ SARS-CoV-2 Specific Furin Cleavage Site (P681R Mutation) Blocker/Inhibitor Screening Kit is a complete set of optimized buffers and reagents designed for screening Blockers of SARS-CoV-2 specific furin cleavage site (FCS) and/or inhibitors of furin and other serine proteases such as human air trypsin (HAT) that may also target the FCS of SARS-CoV-2 in a fast and high throughput format. The kit has the following advantages:

• Colorimetric assay with easy-to-follow steps
• Extremely low background signals
• Simple, accurate, reliable, and consistent assay
• Screening of both SARS-CoV-2 P681R mutated FCS direct blockers or furin and furin-like protease activity inhibitors
• Substrate contains the entire SARS-CoV-2-S1/S2 cleavage sequence
• Uniquely specific for detecting SARS-CoV-2 P681R mutated FCS cleavage and its inhibition

Fig 1. SARS-CoV-2 spike protein cleavage by proprotein convertases and other proteases during entry into cells.

Principles and Procedure

Fig.2. Schematic procedure of the CoviDrop SARS-coV-2 Specific Furin Cleavage Site (Wild type) blocker/Inhibitor Screening Kit

This kit contains all reagents necessary for screening various blockers/inhibitors against SARS-CoV-2-FCS. In this assay, a SARS-CoV-2 specific substrate is tagged with polyhistidine at N-terminal and biotin at C-terminal and bound onto microplate wells through histidine/Ni-NTA at its N-terminal. The cleavage of the substrate at S1/S2 site will remove the S2 part (C-terminal) of the substrate after washing, which causes the decrease in signal generated by avidin/biotin binding after adding Cleavage Detection Solution (CDS). The inhibition of the cleavage by inhibitors will block the reduction of the signals. The signal intensity is measured through an ELISA-like reaction by reading the absorbance in a microplate spectrophotometer at a wavelength of 450 nm. The more the substrate is cleaved, the lower the signal that will be generated. Thus, the PC cleavage activity is inversely proportional to the signal intensity.

Starting Materials

Input materials can be small molecule compounds and other biological molecules that block SARS-CoV-2 specific FSC and/or inhibit furin and serine proteases activity against SARS-CoV-2 specific FCS.