Description
Introduction:
The GENLISA 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 competitiveELISA 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 ofneutralizing-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.
Intended Use:
The GENLISA Human SARS-CoV-2 (2019-nCoV) Spike RBD Quantitative ELISA kit is used as an analytical tool for quantitative estimation of SARS-CoV-2 (2019-nCoV) Spike RBD Antigen in human serum.
Principle:
The method employs sandwich ELISA technique. Monoclonal antibody specific for SARS-CoV-2 (2019-nCoV) Spike RBD is pre-coated onto microwells. Samples and standards are pipetted into microwells and SARS-CoV2 (2019-nCoV) Spike RBD Antigen present in the sample is bound by the immobilized antibody. After incubation the wells are washed and followed by addition of HRP-conjugated Detection anti- SARS-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.
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