Detection of SARS-CoV-2 Receptor-Binding Domain Antibody using a HiBiT-Based Bioreporter

作者： Reza Rezaei1,2, Abera Surendran1,2, Ragunath Singaravelu1,2, Taylor R. Jamieson1,2, Parisa Taklifi3, Joanna Poutou1,2, Taha Azad1,2, Carolina S. Ilkow1,2 1Ottawa Hospital Research Institute, 2Department of Biochemistry, Microbiology and Immunology,University of Ottawa, 3Department of Biotechnology,University of Tehran

简介：

Overview

This protocol outlines a high throughput assay for the rapid and sensitive detection of SARS-CoV-2 antibodies in patient serum samples. The method addresses the need for both speed and sensitivity in serological testing during the COVID-19 pandemic.

Key Study Components

Area of Science

• Immunology
• Virology
• Serological Assays

Background

• COVID-19 has created a global health crisis.
• Current serological assays often compromise either speed or sensitivity.
• There is a need for improved testing methods to detect antibodies.
• The HiBiT-receptor-binding domain protein complex is utilized in this assay.

Purpose of Study

• To develop a novel assay for detecting anti-SARS-CoV-2 antibodies.
• To provide a method that combines both rapid response and high sensitivity.
• To facilitate timely diagnosis and monitoring of COVID-19.

Methods Used

• Preparation of cell culture using Dulbecco's modified eagle medium.
• Incorporation of fetal bovine serum and antibiotics in the culture medium.
• Production of HiBiT-RBD bioreporter for antibody detection.
• High throughput assay for analyzing patient serum samples.

Main Results

• The assay demonstrated both sensitivity and quick response time.
• It allows for the efficient detection of SARS-CoV-2 antibodies.
• Results indicate improved performance over existing serological tests.
• The method is suitable for large-scale testing in clinical settings.

Conclusions

• The developed assay is a valuable tool for COVID-19 research.
• It addresses critical needs in serological testing during the pandemic.
• Future applications may extend beyond SARS-CoV-2 to other pathogens.

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