Simple and ultrastable all-inclusive pullulan tablets for challenging bioassays

Area of Science:

• Biochemistry
• Analytical Chemistry
• Biotechnology

Background:

• Biodetection systems often rely on sensitive enzymes and substrates, limiting their practical use in field or point-of-care settings.
• Current methods require special handling and storage for labile biological components, hindering routine application.
• There is a need for stabilized, user-friendly biodetection formats.

Purpose of the Study:

• To develop a novel, stable, and easy-to-use format for biodetection assays.
• To enhance the shelf-life and operational simplicity of enzyme- and substrate-based detection systems.
• To demonstrate the efficacy of the developed format using a model ATP detection system.

Main Methods:

• Creation of all-inclusive pullulan assay tablets incorporating enzymes and substrates.
• Investigating the factors contributing to enhanced stability within the tablet matrix (restricted molecular motion, oxygen impermeability).
• Evaluating the performance of the tablet-based assay for adenosine triphosphate (ATP) detection using the luciferase-luciferin system.

Main Results:

• The pullulan assay tablets significantly enhance the long-term stability of labile enzymes and organic substrates.
• Tablet formulation restricts molecular motion and prevents oxygen ingress, preserving component activity.
• The assay tablets demonstrate rapid dissolution and ease of use in sample testing.
• Sensitive detection of ATP in biological samples was achieved with the tablet-based assay.
• Assay tablet activity remained stable for over one month at room temperature.

Conclusions:

• The developed pullulan assay tablets offer a simple and effective solution for stabilizing biodetection reagents.
• This innovation greatly improves the feasibility of using sensitive biodetection assays in point-of-care and field environments.
• The tablet format simplifies assay procedures, enhances reagent stability, and maintains high sensitivity for applications like ATP monitoring.