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Activity of InvivoGen's Anti-CoV2RBD Monoclonal Antibodies

Last updated: September 2021

Validation assays

InvivoGen's anti-CoV2RBD monoclonal antibodies (mAbs) have been tested in house to compare their activity. The tests consist of Lucia luciferase-based ELISA and FACS screening assays that have been performed using:

  • RBD-Lucia proteins, a collection of Lucia luciferase-tagged Spike RBD proteins from SARS-CoV-2 variants (including the variants of concern), or  
  • 293-SARS2-S-dfur cells, a series of HEK293-derived cells expressing at the surface the Spike protein of the variants of concern.

 

Below are the data obtained with the following anti-SARS-CoV2 Spike antibodies featuring a mouse IgG2a constant region:

   -  Anti-CoV2RBD-cas : REGN10933-derived mAb
   -  Anti-CoV2RBD-imd : REGN10933-derived mAb
   -  Anti-CoV2RBD-bam : LY-CoV555-derived mAb
   -  Anti-CoV2RBD-ete : LY-CoV016-derived mAb

 

Figure 1: Lucia Luciferase-based ELISA

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The binding capacity of InvivoGen's anti-SARS-CoV2RBD mAbs to a set of Spike variants has been validated using a Lucia luciferase-based ELISA.
Anti-murine IgG F(ab’)2 fragment (2 µg/ml) was coated on an ELISA plate overnight. Anti-CoV2RBD-cas-mIgG2a, Anti-CoV2RBD-imd-mIgG2a, Anti-CoV2RBD-bam-mIgG2a, Anti-CoV2RBD-ete-mIgG2a along with RBD-Lucia proteins (original and V2 to V8; 1 µg/ml) were added and incubated for 2 hours at room temperature. After washing (3x times), binding affinity was assessed by measuring the activity of Lucia luciferase in the supernatant using QUANTI-Luc™. Data are shown as a fold change over no antibody.

 

Figure 2: FACS screening assays

FACS screening assays of Spike variants

InvivoGen's anti-SARS-CoV2RBD mAbs allow the detection of Spike surface expression by mammalian cells for a set of SARS-CoV-2 variants of concern.
Human embryonic kidney (HEK)-293 cells featuring stable cell surface expression of Spike variants were incubated with either Anti-CoV2RBD-cas-mIgG2a, Anti-CoV2RBD-imd-mIgG2a, Anti-CoV2RBD-bam-mIgG2a, or Anti-CoV2RBD-ete-mIgG2a mAbs for 1 hour at 4°C. Cells were then washed and incubated with a PE-conjugated goat anti-murine IgG for 1 hour at 4°C, and surface staining was analyzed by FACS. The grey line indicates the maximal fluorescence intensity for unstained cells.

 

 
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