Split aptamer based sensing platform for adenosine deaminase detection by fluorescence resonance energy transfer

Area of Science:

• Biochemistry
• Analytical Chemistry
• Nanotechnology

Background:

• Adenosine deaminase (ADA) is a crucial enzyme in immune function, and its altered activity is linked to various diseases.
• Accurate and sensitive detection of ADA activity is vital for disease diagnosis and monitoring.
• Existing methods for ADA detection may have limitations in sensitivity, specificity, or sample throughput.

Purpose of the Study:

• To develop a novel, sensitive, and accurate platform for determining adenosine deaminase (ADA) activity.
• To utilize a split aptamer-based fluorescence resonance energy transfer (FRET) system for enzyme detection.
• To validate the platform's performance using biological samples.

Main Methods:

• Construction of a FRET platform using gold nanoclusters (AuNCs) and gold nanoparticles (AuNPs) functionalized with split aptamer fragments.
• Utilizing the interaction of adenosine triphosphate (ATP) with split aptamers to modulate FRET signals.
• Monitoring fluorescence changes upon ADA-catalyzed conversion of ATP to inosine triphosphate (ITP) for enzyme activity determination.

Main Results:

• The FRET platform demonstrated a linear response between fluorescence intensity and ADA concentration from 2-120 U L^-1.
• A low limit of detection (LOD) of 0.72 U L^-1 was achieved, indicating high sensitivity.
• Successful detection of ADA activity in human serum samples confirmed the platform's accuracy and applicability in real-world scenarios.

Conclusions:

• The developed split aptamer-based FRET platform provides a sensitive and reliable method for ADA activity detection.
• This approach offers a valuable tool for clinical diagnostics and biochemical research.
• The platform's design highlights the potential of aptamer-based FRET systems in enzyme activity assays.