SARS-CoV-2 Spike RBD Antigen Quantitative ELISA


The SARS-CoV-2 Spike RBD Antigen Quantitative ELISA for the Quantitative Estimation of SARS-CoV-2 (2019-nCoV) Spike RBD Antigen in human serum. The SARS-CoV-2 Spike RBD Antigen Quantitative ELISA is intended for research use only and not for use in diagnostic or clinical procedures.

SARS-CoV-2 Spike RBD Antigen Quantitative ELISA

The SARS-CoV-2 Spike RBD Antigen Quantitative ELISA is For Research Use Only

Size: 12×8 wells
Standard Range: 0-4000 ng/ml
Incubation Time: 3 hours 20 minutes
Sample Type: Serum
Sample Size: 100 µL
Alternative Name: Coronavirus, SARS-CoV, Covid-19

Assay Principle

The method employs sandwich ELISA technique. Monoclonal antibody specific for SARS-CoV-2 (2019-nCoV) Antibodies to Spike RBD is pre-coated onto microwells. Samples and standards are pipetted into microwells and SARS-CoV-2 (2019-nCoV) Spike RBD Antigen present in the sample are bound by the immobilized antibody. After incubation the wells are washed and followed by addition of HRP-conjugated Detection antiSARS-CoV-2 (2019-nCoV) Spike RBD antibody into each well and incubated to form a complex. After washing microwells in order to remove any non-specific binding, the substrate solution (TMB) is added to microwells and color develops proportionally to the amount of SARS-CoV-2 (2019-nCoV) Spike RBD in the sample. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm.

Sample Preparation and Storage
Specimens should be clear and non-hemolyzed. Samples should be run at a number of dilutions to ensure accurate quantitation.
Blood is taken by venipuncture. Serum is separated after clotting by centrifugation. Repeated freezing and thawing should be avoided. If samples are to be used for several assays, initially aliquot samples and keep at -20°C.
Samples should be diluted 1:1000 (v/v) for optimal recovery, (for example 1 ul sample + 999 ul sample diluent) prior to assay. In cases where matrix interferences is under or over observed, the samples may be diluted with Sample Diluent accordingly.
The samples may be kept at 2 – 8°C for up to three days. For long-term storage please store at -20°C.
Note: Grossly hemolyzed samples are not suitable for use in this assay

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Additional Information

Assay Background

The ELISA kits are used for assessing the specific biomarker in samples analytes which may be serum, plasma and cell culture supernatant as validated with the kit. The kit employs a sandwich ELISA technique which leads to a higher specificity and increased sensitivity compared to conventional competitive ELISA kits which employ only one antibody. The spike (S) glycoprotein of coronaviruses contains protrusions that will only bind to certain receptors on the host cell. Known receptors which bind S1 are ACE2 (angiotensin-converting enzyme 2), DPP4 (dipeptidyl peptidase-4), APN (aminopeptidase N), CEACAM (carcinoembryonic antigen-related cell adhesion molecule 1), Sia (sialic acid), O-ac Sia (O-acetylated sialic acid). The spike is essential for both host specificity and viral infectivity. The term ‘peplomer’ is typically used to refer to a grouping of heterologous proteins on the virus surface that function together. The spike (S) glycoprotein of coronaviruses is known to be essential in the binding of the virus to the host cell at the advent of the infection process. It’s been reported that 2019-nCoV can infect the human respiratory epithelial cells through interaction with the human ACE2 receptor. The spike protein is a large type I transmembrane protein containing two subunits, S1 and S2. S1 mainly contains a receptor binding domain (RBD), which is responsible for recognizing the cell surface receptor. S2 contains basic elements needed for the membrane fusion. The S protein plays key parts in the induction of neutralizing-antibody and T-cell responses, as well as protective immunity.
The main functions for the Spike protein are summarized as:

  • Mediate receptor binding and membrane fusion;
  • Defines the range of the hosts and specificity of the virus;
  • Main component to bind with the neutralizing antibody;
  • Key target for vaccine design;
  • Can be transmitted between different hosts through gene recombination or mutation of the receptor binding domain (RBD), leading to a higher mortality rate.

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