The CoviDrop™ SARS-CoV-2 Spike-ACE2 Binding Inhibitor Screening Fast Kit is a complete set of optimized buffers and reagents designed for screening SARS-CoV-2-ACE2 binding inhibitors using purified SARS-CoV-2 spike protein and ACE2 protein in a fast and high throughput format. The kit has the following advantages:
BackgroundCOVID-19 is an infectious disease caused by SARS-CoV-2, a new member of the same coronavirus family that caused SARS and MERS. Entry of SARS-CoV-2 into human host cells occurs through binding of surface unit S1 of its spike protein to the cell receptor angiotensin-converting enzyme 2 (ACE2). ACE2 is ubiquitous and widely expressed in the lung, heart, kidney, vessels, brain, gut, and testis at different levels. ACE2 is also mostly bound to cell membranes and normally less present in circulation [1]. It was found that increased ACE2 expression in nasal epithelium and lung is associated with higher rates of SARS-CoV-2 infection and severity of COVID-19, respectively [2,3]. A new observation indicates that increased ACE2 concentration in circulation may be associated with increased incidence and fatality rate of COVID-19 [4]. In addition, SARS-CoV-2 invasion degrades ACE2 on cell membranes and may increase soluble ACE2 concentration in circulation [1]. The importance of screening inhibitors of SARS-CoV-2-ACE2 binding is emphasized by the complicated SARS-CoV-2-ACE2 interaction under normal (healthy) and abnormal (pre-existing diseases and viral infection) conditions, the current COVID-19 pandemic, and its possible resurgence.
Fig 1. SARS-Cov-2 binding to ACE2 and entry into cells
Principles and ProcedureThis kit contains all reagents necessary for screening various inhibitors of SARS-CoV-2 spike-ACE2 binding activity or inhibition. In this assay, a SARS-CoV-2 spike protein is stably pre-coated onto microplate wells. His-tagged ACE2 is bound to the coated spike protein in the presence or absence of inhibitors. The amount of the bound ACE2, which is proportional to ACE2 inhibition intensity, is then recognized by the Binding Detection Solution containing anti-His antibody and measured through an ELISA-like reaction by reading the absorbance in a microplate spectrophotometer at a wavelength of 450 nm. The binding activity of ACE2 is proportional to the optical density intensity measured. The more the ACE2 binding is inhibited, the lower the OD intensity is.
Starting MaterialsInput materials can be small molecule compounds, antibodies, and other biological molecules that inhibit or interfere with the binding of SARS-CoV-2 to ACE2.
References: 1. Verdecchia P et al: Eur J Intern Med. 76: 14–20, 2020 2. Bunyavanich S et al: JAMA. Published online May 20, 2020. 3. Sama IZ et al: Eur Heart l, 41: 1810–1817, 2020. 4. Pinto BGG et al: medRxiv. Published online March 27, 2020.