A general chemiluminescence strategy for measuring aptamer-target binding and target concentration

Area of Science:

• Biochemistry
• Analytical Chemistry
• Biotechnology

Background:

• Aptamer-target interactions are crucial in biomedical and analytical applications.
• Measuring aptamer-target binding constants and sites remains a significant challenge.

Purpose of the Study:

• To develop a sensitive, label-free chemiluminescence (CL) strategy for quantifying aptamer-target interactions.
• To establish a general platform for investigating binding constants and sites for diverse targets.
• To characterize ATP binding to a specific aptamer fragment.

Main Methods:

• A label-free chemiluminescence (CL) strategy utilizing N-(aminobutyl)-N-ethylisoluminol functionalized gold colloids.
• Application of the CL method to small molecules, peptides, and proteins.
• Molecular dynamic simulation to analyze ATP-aptamer fragment interactions.

Main Results:

• The CL strategy accurately determines target concentration, binding constant, and binding site.
• The method demonstrates high sensitivity, requiring only nanogram samples.
• It successfully characterized ATP binding to a 14-mer aptamer fragment, including binding constant, site, and conformation.

Conclusions:

• The developed CL strategy provides a sensitive, rapid, and cost-effective tool for studying aptamer-target interactions.
• This platform facilitates aptamer selection and the investigation of biologically important molecular binding.
• The study elucidated specific binding characteristics between ATP and a derived aptamer fragment.