Toward a Point-of-Need Bioluminescence-Based Immunoassay Utilizing a Complete Shelf-Stable Reagent.

Area of Science:

• Biochemistry
• Molecular Biology
• Assay Development

Background:

• Enzyme-linked immunosorbent assays (ELISAs) are widely used but complex for point-of-need testing.
• There is a need for simpler, faster, and sensitive biomolecule detection methods.

Purpose of the Study:

• To develop a bioluminescence-based immunoassay for sensitive and simple biomolecule detection.
• To create a stable, add-and-read assay reagent for clinical diagnostics and research.

Main Methods:

• Utilized a ternary, split-NanoLuc luciferase complementation reporter system.
• Employed directed evolution to optimize reporter stability and solubility.
• Developed formulation optimization for a stable, all-in-one lyophilized reagent.

Main Results:

• Created a shelf-stable, add-and-read assay detection reagent.
• Achieved a robust, first-generation bioluminescence homogenous immunoassay platform.
• Demonstrated one-step, rapid, and sensitive biomolecule quantification in complex human samples.

Conclusions:

• The developed bioluminescence immunoassay offers a sensitive and simple alternative to traditional ELISAs.
• This technology facilitates rapid molecular detection closer to the point-of-need.
• The stable, lyophilized reagent format enhances assay utility in diverse settings.