Journal
BIOSENSORS & BIOELECTRONICS
Volume 121, Issue -, Pages 153-158Publisher
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2018.08.069
Keywords
Telomerase; HeLa cell; pH meter; Bromocresol purple; Urease
Categories
Funding
- National Sciences Foundation of China [21575027, 21575025, 21675028]
- Program for Changjiang Scholars and Innovative Research Team in University [IRT15R11]
- cooperative project of production and study in University of Fujian Province [2018Y4007]
- Professional Foundation of Fujian Province [2018J01685, 2018J01682]
- STS Key Project of Fujian Province [2017T3007]
- Science and Technology Program of Fujian Provincial Bureau of Quality and Technical Supervision [FJQI2014047]
- Foundation for Scholars of Fuzhou University [XRC-1671, XRC-17007]
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This study developed a portable and sensitive biosensor for the detection of telomerase activity extracted from HeLa cells by monitoring urease regulated pH change. In the presence of telomerase, the telomerase substrate (TS) primer was elongated by adding telomeric repeats (TTAGGG)(n) firstly. The TS primer was then immobilized onto streptavidin magnesphere paramagnetic particles (PMPs). The elongated repeat unit can hybridize with biotinylated complementary DNA (cDNA) to specifically capture biotinylated urease onto PMPs. After magnetic separation, the PMPs complex was mixed with urea solution. As urease can catalyze the hydrolysis of urea into ammonia and induce pH increasing of the solution, the activity of telomerase can be detected by pH-sensitive dyes or by a portable pH meter easily. The changed pH has a linear relationship with the logarithm of the number of HeLa cells from 50 to 10,000 and with a detection limit of similar to 20 HeLa cells, which indicated the proposed method have potential application in telomerase-related cancer diagnosis.
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