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SARS-CoV-2 Spike RBD-ACE2 Blocking Antibody Detection ELISA Kit

SARS-CoV-2 Spike RBD-ACE2 Blocking Antibody Detection ELISA Kit #69486

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Supporting Data

REACTIVITY

All

Application Key:

ELISA-ELISA

Species Cross-Reactivity Key:

All-All Species Expected

Product Information

Product Description

The SARS-CoV-2 Spike RBD-ACE2 Blocking Antibody Detection ELISA Kit allows for the detection of antibodies (or small molecules) that block the interaction between the host receptor-binding domain (RBD) of the SARS-CoV-2 spike protein and ACE2. In this assay, SARS-CoV-2 spike RBD protein linked to HRP (RBD-HRP) is pre-incubated with the sample and controls, allowing blocking antibodies present in the sample to bind to the RBD-HRP. These pre-incubated mixtures are then added to microwell plates coated with ACE2 protein. The RBD-HRP is captured by ACE2 in the well to varying degrees depending on the blocking activity present in each sample. The wells are then washed to remove unbound material. HRP substrate, TMB, is added to develop color. The magnitude of optical density for this developed color is inversely proportional to the ability of the sample to block the interaction between SARS-CoV-2 spike RBD and ACE2 proteins. For example, if no blocking activity is present in the sample, the RBD-HRP will be able to bind to ACE2 on the microwell plates, resulting in high signal. Conversely, samples with high blocking activity will prevent the RBD-HRP from binding to ACE2 on the microwell plates, resulting in low signal. This kit is designed to detect blocking antibodies present in human serum/plasma samples. However, this kit may also be used to assess the blocking activity of non-human antibodies and small molecules. In the latter scenarios (for non-human antibodies and small molecules), the user will have to determine the appropriate dilution/concentration of their samples to use, along with running the proper controls.

Protocol

Specificity / Sensitivity

The SARS-CoV-2 Spike RBD-ACE2 Blocking Antibody Detection ELISA Kit detects endogenous levels of antibodies that block the interaction between the SARS-CoV-2 spike RBD (318-541) protein and human ACE2 (18-615). This kit is designed to detect blocking antibodies present in human serum/plasma, however, it may also be used to assess blocking activity of non-human antibodies and small molecules.

Species Reactivity:

All Species Expected

Background

The cause of the COVID-19 pandemic is a novel and highly pathogenic coronavirus, termed SARS-CoV-2 (severe acute respiratory syndrome coronavirus-2). SARS-CoV-2 is a member of the Coronaviridae family of viruses (1). The genome of SARS-CoV-2 is similar to other coronaviruses, and is comprised of four key structural proteins: S, the spike protein, E, the envelope protein, M, the membrane protein, and N, the nucleocapsid protein (2). Coronavirus spike proteins are class I fusion proteins and harbor an ectodomain, a transmembrane domain, and an intracellular tail (3,4). The highly glycosylated ectodomain projects from the viral envelope surface and facilitates attachment and fusion with the host cell plasma membrane. The ectodomain can be further subdivided into host receptor-binding domain (RBD) (S1) and membrane-fusion (S2) subunits, which are produced upon proteolysis by host proteases at S1/S2 and S2’ sites. S1 and S2 subunits remain associated after cleavage and assemble into crown-like homotrimers (2,4). In humans, both SARS-CoV and SARS-CoV-2 spike proteins utilize the angiotensin-converting enzyme 2 (ACE2) protein as a receptor for cellular entry (5-7). Spike protein subunits represent a key antigenic feature of coronavirus virions, and therefore represent an important target of vaccines, novel therapeutic antibodies, and small-molecule inhibitors (8,9).

Database Links

UniProt ID: P0DTC2

Entrez-Gene Id: 43740568

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For Research Use Only. Not For Use In Diagnostic Procedures.

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