期刊
MICROCHIMICA ACTA
卷 184, 期 3, 页码 773-779出版社
SPRINGER WIEN
DOI: 10.1007/s00604-016-2065-3
关键词
Fluorescence; Nanocluster; Enzyme activity assay; TdT; Lymphatic leukemia cells
资金
- National Natural Science Foundation of China [21275096, 21475083]
- Shaanxi Provincial Natural Science Foundation [2013SZS08-Z01]
- Program for Innovative Research Team in Shaanxi Province [2014KCT-28]
The authors report on a simple strategy for sensitive determination of the activity of terminal deoxynucleotidyl transferase (TdT) using copper nanoclusters (CuNCs) as fluorescent probes. TdT-polymerized long chain AT-rich DNA serves as a template for the synthesis of the CuNCs, and TdT activity is detected fluorometrically at excitation/emission wavelengths of 340/570 nm. The protocol relies on the target-triggered formation of dsDNA polymers and in-situ formation of CuNCs. The calibration plot is linear in the 0.7 to 14 U L-1 activity range, with a 60 mU L-1 detection limit (at a signal-to-noise ratio of 3). The protocol was applied to determine TdT activity in acute lymphatic leukemia cells. This approach is selective, simple, convenient and cost-efficient because a complex DNA sequence is not required. In our perception, the method provides a viable new platform for monitoring the activity and inhibition of TdT.
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