Development of a rapid, simple, and sensitive point-of-care technology platform utilizing ternary NanoLuc

Area of Science:

• Biotechnology
• Biochemistry
• Medical Diagnostics

Background:

• Traditional diagnostic tests like ELISAs require complex lab setups and trained personnel, limiting point-of-care applications.
• Point-of-care tests are crucial for rapid medical decision-making and patient care flexibility.
• Developing simple, sensitive, and rapid diagnostic methods for point-of-care use remains a significant challenge.

Purpose of the Study:

• To develop a homogeneous, bioluminescent-based, split reporter platform for simple, sensitive, and rapid analyte detection at the point-of-care.
• To create a shelf-stable, all-in-one, add-and-read system for complex clinical sample matrices.
• To demonstrate the platform's modularity and utility using SARS-CoV-2 detection models.

Main Methods:

• Utilized an optimized ternary, split-NanoLuc luciferase reporter system with analyte-specific affinity reagents.
• Engineered reporter peptides for proximity-induced complementation upon analyte binding, generating a bioluminescent signal.
• Incorporated lyophilization for a stabilized, shelf-stable reporter system and substrate in an all-in-one format.

Main Results:

• Demonstrated a simple, sensitive, and rapid method for analyte detection in complex sample matrices.
• Successfully developed SARS-CoV-2 N-antigen and anti-SARS-CoV-2 antibody detection assays using the platform.
• Showcased flexible assay formatting with chemically conjugated or genetically fused affinity reagents.

Conclusions:

• The developed bioluminescent split reporter platform offers a standardized and simple method for rapid, robust, and sensitive analyte detection.
• This technology is ideal for research, clinical laboratory, and point-of-care applications, requiring only a handheld luminometer.
• The platform's modularity and ease of use facilitate the development of diverse diagnostic assays.