A Label-Free Fluorescent Assay for the Rapid and Sensitive Detection of Adenosine Deaminase Activity and Inhibition.

摘要: Adenosine deaminase (ADA), able to catalyze the irreversible deamination of adenosine into inosine, can be found in almost all tissues and plays an important role several diseases. In this work, we developed a label-free fluorescence method for detection activity inhibition. presence ADA, ATP has been shown hydrolyzed. The aptamer was form G-quadruplex/thioflavin T (ThT) complex with ThT exhibited obvious signal. However, could bind low signal because absence ADA. This assay showed high sensitivity ADA limit 1 U/L based on SNR 3 got good linear relationship within range 1⁻100 R² = 0.9909. LOD is lower than cutoff value (4 U/L) clinical requirement more sensitive most reported methods. technique selectivity against hoGG I, UDG, RNase H λexo. Moreover, strategy successfully applied assaying inhibition using erythro-9-(2-hydroxy-3-nonyl) adenine (EHNA) and, as such, demonstrated great potential future use diagnosis ADA-relevant diseases, particularly advanced drug development.