Enzyme-based digital bioassay technology - key strategies and future perspectives

Area of Science:

• Bioanalytical Chemistry
• Biotechnology
• Molecular Diagnostics

Background:

• Digital bioassays utilize single-molecule reactions for highly sensitive and quantitative biomolecule detection.
• This approach involves compartmentalizing molecules at the single-molecule level within microreactors.
• The core principle is counting microreactors that exhibit positive signals, indicating the presence of target molecules.

Purpose of the Study:

• To provide a comprehensive overview of digital bioassay technologies.
• To highlight the evolution and diverse applications of digital bioassays, including digital ELISA.
• To discuss future perspectives and emerging fields driven by digital bioassay advancements.

Main Methods:

• Review of existing literature on digital bioassays and single-molecule detection.
• Categorization of digital bioassays based on their principles and applications.
• Discussion of microreactor technologies enabling advanced digital bioassays.

Main Results:

• Digital bioassays, including digital ELISA, have shown significant growth since the 2000s.
• Applications have expanded from enzymes to complex systems like membrane transporters and cell-free gene expression.
• Development of multiplex and multiparametric digital bioassays allows for precise assessment of sample heterogeneities.

Conclusions:

• Digital bioassays represent a powerful and versatile platform for sensitive biomolecule detection.
• Ongoing advancements in microreactor technology are continuously expanding the scope of digital bioassay applications.
• The future holds promise for novel digital bioassays addressing complex biological questions and diagnostics.