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Published on: August 12, 2021
Using Split Luminescent Biosensors for SARS-CoV-2 Antibody Detection in Serum, Plasma, and Blood Samples
Susanna K Elledge1, Ian Eigl1, Maira Phelps2
1Department of Pharmaceutical Chemistry, University of California, San Francisco, California.
Insights
A novel split nanoluciferase assay enables rapid, quantitative detection of SARS-CoV-2 antibodies in various settings. This assay accurately tracks immunity over time, offering a significant advancement for COVID-19 monitoring and future viral immunity evaluations.
Area of Science:
- Biotechnology
- Immunology
- Virology
Background:
- Accurate antibody detection is crucial for assessing viral immunity, especially for COVID-19.
- Existing serology assays like ELISA are time-consuming and lab-dependent, while lateral flow assays (LFAs) are qualitative and lack longitudinal tracking capabilities.
- A need exists for rapid, simple, and quantitative antibody detection methods to monitor immunity over time.
Purpose of the Study:
- To develop a novel, rapid, and quantitative assay for detecting SARS-CoV-2 antibodies.
- To enable accurate tracking of antibody levels in patients over time.
- To provide a versatile assay adaptable to different settings, from laboratories to point-of-care.
Main Methods:
- Developed a split nanoluciferase assay utilizing SARS-CoV-2 spike or nucleocapsid proteins.
- Enabled luminescent-based detection of specific antibody binding in serum, plasma, and whole blood.
- Validated the assay for medium-throughput (plate-based), high-throughput (robotic), and point-of-care (handheld luminometer) applications.
Main Results:
- The split-luciferase assay provides simple, rapid, and quantitative detection of SARS-CoV-2 antibodies.
- The assay is highly adaptable for various throughput scales and settings.
- Demonstrated successful implementation in laboratory and point-of-care environments.
Conclusions:
- The developed split-luciferase assay offers a versatile and efficient method for SARS-CoV-2 antibody detection.
- This assay simplifies and reduces assay time, enhancing accessibility in clinical and laboratory settings.
- The technology facilitates antibody detection in non-laboratory, point-of-care environments, improving public health surveillance capabilities.
Abstract:
Antibody detection assays are essential for evaluating immunity of individuals against a given virus, and this has been particularly relevant during the COVID-19 pandemic. Current serology assays either require a laboratory setting and take >1 hr (i.e., enzyme-linked immunosorbent assay [ELISA]) or are rapid but only qualitative in nature and cannot accurately track antibody levels over time (i.e., lateral flow assay [LFA]). Therefore, there is a need for development of a rapid and simple but also quantitative assay that can evaluate antibody levels in patients accurately over time. We have developed an assay that uses a split nanoluciferase fused to the spike or nucleocapsid proteins of the SARS-CoV-2 virus to enable luminescent-based detection of spike- or nucleocapsid-binding antibodies in serum, plasma, and whole blood samples. The resulting approach is simple, rapid, and quantitative and is highly amenable to low-/medium-throughput scale using plate-based assays, high-throughput scale using robotics, and point-of-care applications. In this article, we describe how to perform the assay in a laboratory setting using a plate reader or liquid-handling robotics and in a point-of-care setting using a handheld, battery-powered luminometer. Together, these assays allow antibody detection to be easily performed in multiple settings by simplifying and reducing assay time in a laboratory or clinical environment and by allowing for antibody detection in point-of-care, nonlaboratory settings. © 2022 Wiley Periodicals LLC. Basic Protocol: SARS-CoV-2 antibody detection using the split-luciferase assay on a medium-throughput scale with a laboratory luminometer Alternate Protocol 1: High-throughput-based protocol for SARS-CoV-2 antibody detection using a robotic platform Alternate Protocol 2: Point-of-care-based protocol for SARS-CoV-2 antibody detection using a handheld luminometer Support Protocol: Determining positive/negative cutoffs for test samples and standardizing the assay between days.
