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