Label-free DNA hairpin probe for real-time monitoring of alkaline phosphatase activity. Issue 25 (13th June 2016)

• Record Type: Journal Article
• Authors: Du, Junyan; Xiong, Leqin; Ma, Changbei; Liu, Haisheng; Wang, Jun; Wang, Kemin
• Is Part Of: Analytical methods. Volume 8:Issue 25(2016)
• Page Start: 5095
• Page End: 5100
• Publication Date: 2016-06-13
• Subjects: Chemistry, Analytic -- Periodicals; Analytical biochemistry -- Periodicals; Chemical laboratories -- Standards -- Periodicals; 543.1905
• DOI: 10.1039/c6ay00989a
• Languages: English
• ISSNs: 1759-9660

Abstract

Abstract : A simple fluorescence-based biosensor for the label-free, real-time detection of alkaline phosphatase activity is reported. Abstract : We have developed a new methodology for the label-free detection of alkaline phosphatase (ALP) activity based on a DNA hairpin and thioflavin T (ThT) probes. In the presence of ALP, the 3′-phosphate of the primer DNA is dephosphorylated, triggering a primer extension reaction to open the hairpin probe and leading to release of the G-quartets. These then bind to ThT to form ThT/G-quadruplexes with obvious fluorescence generation. The method is highly sensitive, and the ALP detection limit was estimated to be 0.001 U mL −1, which is superior or comparable to those of reported assays. The assay avoids complicated modifications or chemical labeling, making it simple and cost-effective. Furthermore, the inhibition of ALP activity by the inhibitor theophylline is demonstrated, indicating the potential for the discovery and characterization of ALP-targeted drug candidates in pharmaceutical development.