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Lumit™ SARS-CoV-2 Spike RBD: hACE2 Immunoassay

No-Wash Detection of Spike (RBD) and Human ACE2 Interaction

  • Detect neutralizing antibodies, peptides, or small molecule inhibitors
  • Linear response in the picomolar to ~2nM range of SARS-CoV-2 RBD and hACE2 proteins
  • No wash, transfer or immobilization steps.
  • Available as early access material; please contact us for more information.

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This product is discontinued
Lumit™ SARS-CoV-2 Spike RBD: hACE2 Immunoassay

Scalable Assay to Monitor Viral:ACE2 Interaction

The Lumit™ SARS-CoV 2 Spike RBD: hACE2 Immunoassay* is a homogeneous bioluminescent assay that measures levels of interaction between SARS-CoV-2 Spike protein receptor binding domain (RBD) and human angiotensin-converting enzyme 2 (ACE2).

NanoBiT® subunits are conjugated to a pair of secondary antibodies against two different species (anti-rabbit and anti-mouse antibodies). To detect interaction between RBD and ACE2, a rabbit Fc domain-tagged RBD and a mouse Fc domain-tagged ACE2 are mixed together with SmBiT- and LgBiT-conjugated secondary antibodies (Lumit™ Anti-Mouse Ab-LgBiT and Lumit™ Anti-Rabbit Ab-SmBiT). Interaction of RBD and ACE2, along with binding of the Lumit™ secondary antibodies to their corresponding epitopes on the Fc-domains, brings NanoBiT® subunits into proximity to form an active NanoLuc® luciferase enzyme that generates light in proportion to the amount of target protein.

*For Research Use Only. Not for Use in Diagnostic Procedures.

Assay Principle

 SARS-CoV 2 Spike RBD: hACE2 Immunoassay principle

Lumit™ SARS-CoV 2 Spike RBD: hACE2 Immunoassay principle. Detection of RBD-rabbit Fc:hACE2-mouse Fc interaction is performed by Lumit™ secondary antibodies (Lumit™ Anti-Mouse Ab-LgBiT and Lumit™ Anti-Rabbit Ab-SmBiT). Panel A. Binding of the antibodies to the proteins promotes the reconstitution of NanoLuc® luciferase, generating bioluminescence. Panel B. The presence of a molecule that disrupts the RBD-hACE2 interaction reduces the bioluminescent signal.

Surrogate Neutralization Assay

Neutralization of RBD ACE2 Interaction

Detection of neutralizing effects of patient serum samples with the Lumit™ SARS-CoV 2 Spike RBD: hACE2 Immunoassay. PCR-positive (n = 40) and pre-pandemic (n = 43) patient serum samples were incubated with 1.5nM of each of SARS-CoV-2 RBD (Rabbit Fc), hACE2 (Mouse Fc) and Lumit™ Antibody Mix. After a 1 hour incubation at 23°C, 12.5μl Lumit™ Detection Reagent was added, followed by incubation at room temperature for 30 minutes. Luminescence was recorded using a GloMax® Discover Microplate Luminometer (Cat.# GM3000). Net luminescence values of all samples were converted to percentage PPI activity using the “Lumit Reagents Only” and “Pre-COVID19 negative serum” controls as 0% and 100%, respectively. Sample percent neutralization was calculated as [100 – % PPI activity]. The dotted line represents the cutoff of at 30% neutralization/inhibition.

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