Aptamer binding assays and molecular interaction studies using fluorescence anisotropy - A review

Area of Science:

• Biochemistry
• Molecular Biology
• Analytical Chemistry

Background:

• Nucleic acid aptamers are versatile recognition elements for specific molecular targets.
• Fluorescence anisotropy (FA) and fluorescence polarization (FP) are sensitive biophysical techniques for detecting molecular interactions.

Purpose of the Study:

• To review recent advancements in aptamer-based affinity binding assays utilizing FA/FP techniques.
• To highlight the application of these methods in studying molecular interactions and identifying binding sites.

Main Methods:

• Review of studies employing aptamers labeled with fluorophores for FA/FP detection.
• Analysis of how aptamer binding-induced changes in structure, volume, and fluorophore rotation affect FA/FP signals.
• Summarization of studies in homogeneous solutions and separation techniques like capillary electrophoresis.

Main Results:

• Aptamer-based FA/FP assays effectively detect molecular interactions by measuring changes in fluorescence signals.
• These techniques provide insights into binding affinity and precise mapping of binding sites at the nucleotide level.
• Benefits include ease of fluorophore labeling, aptamer structural changes, and adsorption onto nanomaterials.

Conclusions:

• Aptamer-FA/FP assays are powerful tools for characterizing molecular interactions and binding sites.
• The ability to map binding at the single nucleotide level represents a significant advantage for future research directions.