期刊
MICROCHIMICA ACTA
卷 185, 期 2, 页码 -出版社
SPRINGER WIEN
DOI: 10.1007/s00604-017-2628-y
关键词
Pyrophosphate; ABTS; Hemin; Enzyme activity assay; ALP inhibitor
资金
- National Natural Science Foundation of China [21205142]
- Research Innovation Program for Graduates of Central South University [2016zzts580, 2017zzts347]
A colorimetric detection scheme is introduced for the determination of alkaline phosphatase (ALP) activity based on Cu(II)-modulated G-quadruplex-based DNAzymes. It is exploiting the strong affinity of Cu(II) for pyrophosphate (PPi) upon which the cofactor PPi is trapped by Cu(II). Hence, the activity of the DNAzyme is inhibited. ALP catalyzes the hydrolysis of PPi, causing the release of Cu(II). DNAzyme, in turn, is activated and catalyzes the cleavage of the DNA probe substrate. The released G-rich sequence folds into the G-quadruplex, which can bind hemin and catalyze the oxidation of 2,2'-azinobis (3-ethylbenzothiozoline)-6-sulfonate (ABTS), and this leads to an increase in absorbance at 420 nm. Absorbance increases linearly with increasing ALP activity in 0.07 to 300 U.L-1 range, with a 70 mU.L-1 detection limit. The method was applied in ALP inhibition tests and to the determination of ALP activity in spiked serum samples where it gave satisfactory results.
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