Delta Variant - Luciferase-tagged RBD protein

Delta | B.1.617.2 | RBD-Lucia

ABOUT

Recombinant RBD fusion protein (B.1.617.2 variant - Indian origin) for ELISA & LIPS

RBD-LuciaV8 (B.1.617.2) (~52 kDa) is a soluble fusion protein composed of the Spike Receptor Binding Domain (RBD) from the SARS-CoV-2 Delta variant (B.1.617.2) fused to a C‑terminal Lucia luciferase tag. RBD-LuciaV8 (B.1.617.2) has been specifically designed to assess the binding affinity of anti-Spike antibodies using either ELISA or LIPS (luciferase immunoprecipitation systems) assays [1-3].

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SARS-CoV-2 Spike RBD

RBD-LuciaV8 (B.1.617.2) contains the Spike RBD domain, including the receptor-binding motif (RBM) from the SARS-CoV-2 Delta variant, first reported in India in October 2020 [4]. This variant is classified as a member of Clade 21A/ B.1.617.2 lineage (Nextstrain/Pango lineage classification). It is characterized by the presence of two key mutations within the Spike RBD coding region, which are of concern [4-5].

  • L452R, T478K

More details Learn more about the emerging SARS-CoV-2 variants around the world

Applications

Luciferase-tagged RBD proteins are ideal for studying the binding of anti-spike monoclonal antibodies (mAbs) by solid-phase ELISA and/or solution‑phase LIPS assays, as well as anti‑spike polyclonal antibodies in the sera of recovered COVID‑19 patients and/or vaccinees by LIPS [1-3].

  • ELISA: the Lucia luciferase tag provides a larger dynamic range than the commonly used HRP detection.
  • LIPS: for the detection of antibodies, against both linear and conformational epitopes.

RBD‑LuciaV8 (B.1.617.2) has been generated by recombinant DNA technology, produced in CHO cells, and purified by IMAC (Immobilized Metal Affinity Chromatography) using a C‑terminal histidine tag. Protein size and purity (>90%) have been validated by SDS‑PAGE and the absence of endotoxin contamination has been confirmed using cellular assays. 

References:

1. Burbelo, P.D. et al. 2010. Antibody-profiling technologies for studying humoral responses to infectious agents. Expert Rev Vaccines 9, 567-578.
2. Haljasmagi, L. et al. 2020. LIPS method for the detection of SARS-CoV-2 antibodies to spike and nucleocapsid proteins. Eur J Immunol 50, 1234-1236.
3. Liang, Y. et al. 2021. A luciferase immunosorbent assay for quantitative detection of IgG antibodies against SARS-CoV-2 nucleoprotein. J Virol Methods 292, 114141.
4. Davis, C. et al. 2021. Reduced neutralisation of the Delta (B.1.617.2) SARS-CoV-2 variant of concern following vaccination. medRxiv doi:10.1101/2021.06.23.21259327.
5. Centers for Disease Control and Prevention. SARS-CoV-2 Variant Classifications and Definitions. Retrieved 07 July 2021.

All InvivoGen products are for internal research use only, and not for human or veterinary use.

RBD-LuciaV8 (B.1.617.2) fusion protein for ELISA & LIPS
RBD-LuciaV8 (B.1.617.2) fusion protein for ELISA & LIPS

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SPECIFICATIONS

Specifications

Accession number

EPI_ISL_2356230

Tag
His6, C-ter
Purification
HisTrap affinity chromatography
Appearance (form)
Lyophilized powder
Appearance (color)
White
Tested applications

ELISA, LIPS

Quality control

Each lot is functionally tested and validated.

CONTENTS

Contents

  • Product: 
    RBD-LuciaV8 (B.1.617.2)
  • Cat code: 
    rbd-lucia-v8
  • Quantity: 
    50 µg
Includes:
  • 1.5 ml of endotoxin-free water
  • 1 tube of QUANTI-Luc™ 4 Reagent, a Lucia luciferase detection reagent (sufficient to prepare 25 ml)

Shipping & Storage

  • Shipping method:  Room temperature

    • Storage:

    • -20°C

    Caution:

    • Avoid repeated freeze-thaw cycles

Details

RBD-Lucia in ELISA

RBD-Lucia proteins can be used in a luciferase-based ELISA. Unlike a conventional ELISA, the plate is coated overnight with an Anti-human IgG F(ab')2 fragment. Upon addition of anti-spike monoclonal antibodies (mAb), they will bind to this 'capture' fragment through their Fc region, and RBD-Lucia will bind to the variable region. The luciferase activity is then used to assess the mAb binding affinity to the Spike RBD.

RBD-Lucia in LIPS

Currently, to perform a LIPS assay, soluble crude cell lysates or culture media of the luciferase tagged recombinant protein are extracted from transfected cells and directly used for the assay. InvivoGen's RBD-Lucia proteins streamline the protocol even further. Simply add the RBD-Lucia protein to either anti-spike mAbs or to anti‑spike polyclonal antibodies in the sera of recovered COVID‑19 patients and/or a vaccinee. Following this, antibody-protein complexes are purified using Protein A beads. Quantification of either binding affinity (mAb) and/or antibody levels (sera) is easily determined by assessing the Lucia luciferase activity.

DOCUMENTS

Documents

RBD-LuciaV8 (B.1.617.2)

Technical Data Sheet

Validation Data Sheet

Safety Data Sheet

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Certificate of analysis

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