Bioreporter Assay: A Sensitive Technique using a Bioluminescent Reporter System to Detect SARS-CoV-2 Antibodies

Published: April 30, 2023

Abstract

Source: Rezaei, R. et al., Detection of SARS-CoV-2 Receptor-Binding Domain Antibody using a HiBiT-Based Bioreporter. J. Vis. Exp. (2021)

In this video, we show the HiBit-RBD bioreporter system-based assay for detecting SARS-CoV-2 neutralizing antibodies. This system targets the viral RBD domain by using a bioluminescent reporter system containing HiBiT fused to the RBD domain.

Protocol

1. Detecting anti-RBD antibody with a fast and sensitive assay HiBiT-RBD antibody detection assay Prepare the HiBiT-RBD bioreporter. NOTE: It is recommended to use the supernatant for the following assay as it is simpler to collect and contains the mature glycosylated version of the protein. Moreover, the lysate has several other proteins that could interfere with the HiBiT-RBD-antibody interaction. Combine 50 µL of the HiBiT-RBD-containing supernatant with…

Disclosures

The authors have nothing to disclose.

Materials

Bio-Plex Handheld Magnetic Washer Bio-Rad 171020100 Device
Dual-Glo luciferase Assay System Promega E2940 100 mL kit
LgBiT Promega N3030 Assay kit
Pierce Protein G Magnetic Beads Thermo Fisher Scientific 88848 Assay
SARS-CoV-2 (2019-nCoV) Spike Neutralizing Antibody, Mouse Mab SinoBiological 40592-MM57 Assay
Synergy Mx Microplate Reader BioTek 96-well plate reader luminometer

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Cite This Article
Bioreporter Assay: A Sensitive Technique using a Bioluminescent Reporter System to Detect SARS-CoV-2 Antibodies. J. Vis. Exp. (Pending Publication), e21226, doi: (2023).

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