Related Experiment Videos
Nucleic acid-controlled quantum dots aggregation: A label-free fluorescence turn-on strategy for alkaline phosphatase
Zhenzhen Hu1, Jian Chen1, Yongxin Li1
1State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, PR China; University of Chinese Academy of Sciences, Beijing 100049, PR China.
Abstract:
Based on the controlled aggregation of quantum dots (QDs), a valid, reliable, and label-free fluorescence turn-on strategy is established for the detection of alkaline phosphatase activity. The aqueous solution of anionic QDs exhibits intense fluorescence. However, the addition of cationic polymer (poly-1) significantly quenched the fluorescence of the QDs via their induced aggregation. While short 3'-phosphorylated DNA (DNA-P) could not be extended by terminal deoxynucleotidyl transferase (TdT) and therefore, fluorescence of the QDs was recovered negligibly. The effective elimination of phosphate group of DNA-P in the presence of alkaline phosphatase (ALP) produced 3'-OH termini and the resulting DNA could be sufficiently extended by TdT. The presence of greater binding strength between the elongated DNA and poly-1 is very crucial to compete with the poly-1/QDs aggregates and release the QDs. Turned-on fluorescence emission is observed due to the efficient release of the QDs. A novel strategy for alkaline phosphatase detection is therefore established. Our method is quite sensitive and selective, as low as 0.1 mU/mL ALP can be easily detected.