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SARS-CoV-2 HiBiT-PsVLP Bioassay

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Measure Potency and Stability of Inhibitors that Bind and Block Viral Entry into Target Cells

  • Safe: No virus generation; suited for BSL-1/2 environments
  • Simple: Ready-to-use kit format; just add, mix and read
  • Fast: Get data in hours instead of days
  • Flexible: Adaptable for multiple viruses
  • Early access material; learn more

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SARS-CoV-2 HiBiT-PsVLP Bioassay

Convenient Bioassay, Quality Results

Neutralizing antibodies, whether generated through vaccination or produced as monoclonal antibodies, are pivotal tools for stopping viral infections. Determining the neutralizing capability of biologic molecules is a critical parameter.

Current methods, such as PRNT, sVNT or PNA, can assess the ability of antibodies or small molecules to neutralize viruses. However, current methods are either labor-intensive, slow, highly variable or difficult to implement due to biosafety requirements. As a result, these assays are difficult to establish in a quality-controlled setting.

Virus-like particles (VLPs) are virus-derived structures made up of one or more different molecules that can self-assemble. They mimic the size and shape of a viral particle but lack the genetic material necessary for infectivity. By targeting VLPs to specific cell types through pseudotyping or by incorporating attachment proteins from the virus of interest (such as the Spike protein of SARS-CoV-2), one can effectively assess viral attachment and entry into target cells, thereby determining the level of neutralization.

The SARS-CoV-2 HiBiT-PsVLP Bioassay is a bioluminescent reporter cell-based assay used to measure the neutralization capacity of ligands or antibodies that bind and block viral entry. This simple, convenient and safe bioassay overcomes many limitations of the existing methods for determining neutralizing activity.

Bioassay Components and Product Formats

The SARS-CoV-2 HiBiT-PsVLP Bioassay consists of two primary components:

  • SARS-CoV-2 HiBiT-PsVLP: HiBiT-tagged VLPs pseudotyped with SARS-CoV-2 Spike protein
  • SARS-CoV-2 HEK 293T Target Cells: HEK 293T cells engineered with viral entry proteins ACE2 and TMPRSS2, and intracellular LgBiT protein

Complete Kits:

  • Include SARS-CoV-2 HiBiT-PsVLP, SARS-CoV-2 HEK 293T Cells, cell culture media and assay reagents
  • Recommended for routine use with customer-defined antibody or small-molecule inhibitor
  • Available in 1X and 5X sizes

Target Kits:

  • Include SARS-CoV-2 HEK 293T Target Cells, cell culture media and assay reagents
  • Recommended for customers looking to expand testing against SARS-CoV-2 variants
  • Available in 1X, 5X and CPM formats

HiBiT-PsVLP Kits:

  • Include SARS-CoV-2 HiBiT-PsVLPs
  • Recommended for customers looking to expand testing against SARS-CoV-2 variants
  • Available in 1X and 5X sizes for the following viral variants:

When the SARS-CoV-2 HiBiT-PsVLP and SARS-CoV-2 HEK 293T Cells are incubated together, the PsVLPs bind to target cells via the Spike/ACE2 interaction and undergo membrane fusion. This process results in release of HiBiT into the target cells, where it binds to LgBiT to generate a luminescent signal when substrate is present. However, in the presence of inhibitors or neutralizing mAbs (NmAb) of SARS-CoV-2 entry, the entry and fusion processes of PsVLPs are blocked, thereby preventing HiBiT release and luminescent signal.

Workflow

SARS-CoV-2 HiBiT-PsVLP Bioassay Workflow

Standardized Reagents Improve Results

  • Thaw-and-use format provides significant time and labor savings over traditional methods, like PRNT, while reducing variability.
  • Easily amenable to high-throughput workflows.

Bioassay Principle

The SARS-CoV-2 HiBiT-PsVLP Bioassay utilizes pseudotyped virus-like particles (PsVLPs) to assess the effectiveness of small-molecule inhibitors and neutralizing antibodies in preventing viral entry into host cells. The assay uses innovative HiBiT bioluminescence technology to tag PsVLPs that contain the Spike protein on their surface. The PsVLPs are noninfectious because they contain no viral genetic material.

Representation of the SARS-CoV-2 HiBiT-PsVLP Bioassay

Representation of the SARS-CoV-2 HiBiT-PsVLP Bioassay.

Bioassay Performance Data

HiBiT-PsVLP Assay recapitulates differing sensitivity of SARS-CoV-2 variants to neutralization by therapeutic mAbs
HiBiT-PsVLP Assay recapitulates differing sensitivity of SARS-CoV-2 variants to neutralization by therapeutic mAbs
HiBiT-PsVLP Assay recapitulates differing sensitivity of SARS-CoV-2 variants to neutralization by therapeutic mAbs
HiBiT-PsVLP Assay recapitulates differing sensitivity of SARS-CoV-2 variants to neutralization by therapeutic mAbs

HiBiT-PsVLP Bioassay recapitulates differing sensitivity of SARS-CoV-2 variants to neutralization by therapeutic mAbs. SARS-CoV-2 LgBiT/Target Cells were incubated with various SARS-CoV-2 Spike neutralizing antibodies, and each of the SARS-CoV-2 S HiBiT-PsVLPs carried different spike variants (G614, Alpha, Beta, Gamma and Delta). After 3 hours of incubation, Nano-Glo® Live Cell Reagent was added and luminescence was determined in a GloMax® Discover Plate Reader. Imdevimab (Panel B) maintains neutralizing activity against all variants, while the other three NmAbs tested showed varying degrees of neutralizing activity against different variants (Panels A, C and D).


Patents and Disclaimers

Materials may be covered by pending or issued patents or may have certain limitations. Information is available upon request and will be included on applicable quotes.

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