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 blocking ELISA technique which mimics the virus neutralization process. SARS-CoV-2-neutralizing antibodies primarily target the trimeric spike (S) glycoproteins on the viral surface that mediate entry into host cells. The S protein has two functional subunits that mediate cell attachment (the S1 subunit, existing of four core domains S1A through S1D) and fusion of the viral and cellular membrane (the S2 subunit).Potent neutralizing antibodies often target the receptor interaction site in S1, disabling receptor interactions. The spike proteins of SARS-CoV-2 commonly bind to the human angiotensin coverting enzyme 2 (ACE2) protein as a host receptor through their S1B domain. Receptor interaction is known to trigger irreversible conformational changes in coronavirus spike proteins enabling membrane fusion. SARS-CoV-2 initiates an immune response, which leads to the production of antibodies. These neutralizing antibodies provide protection against future infection from SARS-CoV-2 (Variant B.1.1.7), as they remain in the circulatory system for months to years post infection. SARS-CoV-2 keeps evolving by continual mutation which enables the virus to evade vaccines and immune systems. Some of the mutations, the U.K. variant B.1.1.7, the Brazil variant P.1, the South Africa variant B.1.351 and the Indian variant B.1.617, may have allowed the virus to escape from neutralizing antibodies. The combination of N501Y substitution is present in B.1.1.7 which was first reported in United Kingdom. This Neutralizing Kit works on using Recombinant SARS-CoV-2 (2019-nCoV) Spike RBD (N501Y) Protein and a Neutralizing Antibody sensitive to the variant as a Standard/Calibrator to measure the effective inhibition %.
Intended Use:
The GENLISA SARS-CoV-2 (Covid-19) Alpha Neutralizing Antibody (B.1.1.7) ELISA kit is used as an analytical tool for the qualitative and quantitative detection all types of neutralizing antibodies against SARSCoV-2 RBD (N501Y) in serum or plasma.
Principle:
The method employs sandwich ELISA technique. The protein-protein interaction between HRP-RBD and hACE2 can be blocked by neutralizing antibodies against SARS-CoV-2 RBD (N501Y). Samples and controls are pipetted in a blank microtitre plate and incubated with HRP conjugated human SARSCoV-2 RBD (N501Y) protein. The antibodies to SARS-CoV-2 (N501Y) present in the samples and controls bind to the SARS-CoV-2 RBD (N501Y) protein to form a complex. This solution of bound and unbound antibodies to SARS-CoV-2 Variant B.1.1.7 is then pipetted into human ACE2 coated microplate. After washing to remove the bound complex of Anti-SARS-CoV-2 Variant B.1.1.7 and HRP conjugated SARS-CoV-2 RBD (N501Y), the substrate solution (TMB) is added to the microwells. Post incubation, color develops proportionally to the amount of unbound Anti-SARS-CoV-2 (Covid-19) Variant B.1.1.7 present in the sample. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm.
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