Principle of the Assay
This assay employs a competitive ELISA format and is uniquely suitable for rapid high-throughput screening of the Anti-SARS-CoV-2 neutralizing antibody in serum or cell culture supernatants. The ACE2 receptor protein has been pre-coated onto a microplate. Controls or samples are pipetted into the wells, and then a horseradish peroxidase-conjugated recombinant SARS-CoV-2 RBD fragment (RBD-HRP) is added to the wells together. The protein-protein interaction between RBD-HRP and ACE2 can be blocked by neutralizing antibodies against SARS-CoV-2 RBD. Following incubation and wash steps, a substrate is added. A colored product is formed in proportion to the amount of SARS-CoV-2 Neutralization Antibody present in the sample. The reaction is terminated by the addition of acid and absorbance is measured at 450nm. The positive control is prepared from SARS-CoV-2 Neutralization Antibody (Mouse IgG) to assure the validity of the results.
Coronaviruses are enveloped viruses with a positive-sense RNA genome and with a nucleocapsid of helical symmetry. SARS-CoV-2 has several structural proteins including spike (S), envelope (E), membrane (M), and nucleocapsid (N). The spike protein (S) contains a receptor-binding domain (RBD), which is responsible for recognizing the cell surface receptor, angiotensin-converting enzyme-2 (ACE2). It is found that the RBD of the SARS-CoV-2 S protein strongly interacts with the human ACE2 receptor leading to endocytosis into the host cells of the deep lung and viral replication. The subpopulation of the binding antibodies that can block cellular infiltration and replication of the virus are named neutralizing antibodies. This kit applies to the screening of the SARS-CoV-2 neutralization antibody.
|Kit Components||96 Wells Quantity/Size|
|Aluminum pouches with a Microwell Plate coated with Ace2 receptor protein (8×12)
|Positive Control||2 vials|
|Negative Control||2 vials|
|Recombinant SARS-CoV-2 RBD fragment conjugated to horseradish peroxidase (HRP)
|Wash Buffer Concentrate 20x (PBS with 1% Tween-20)||1 bottle|
|Substrate Solution||1 bottle|
|Stop Solution||1 bottle|
|Adhesive Films||4 pieces|
|Product insert||1 copy|
|Storage||Store at 2 - 8°C|
|REPEATABILITY||The coefficient of variation of both intra-assay and inter-assay was less than 10%.|
|SPECIFICITY||This assay is specific to SARS-CoV-2 neutralizing antibodies.|