Label-free electrochemiluminescence biosensor for ultrasensitive detection of telomerase activity in HeLa cells based on extension reaction and intercalation of Ru(phen)3 (2.).
Telomerase is one of the most common markers of human malignant tumors, such as uterine, stomach, esophageal, breast, colorectal, laryngeal squamous cell, thyroid, bladder, and so on. It is necessary to develop some sensitive but convenient detection methods for telomerase activity determination. In this study, a label-free and ultrasensitive electrochemiluminescence (ECL) biosensor has been fabricated to detect the activity of telomerase extracted from HeLa cells. Thiolated telomerase substrate (TS) primer was immobilized on the gold electrode surface through gold-sulfur (Au-S) interaction and then elongated by telomerase specifically. Then, it was hybridized with complementary DNA to form double-stranded DNA (dsDNA) fragments on the electrode surface, and Ru(phen)3 (2+) has been intercalated into the dsDNA grooves to act as the ECL probe. The enhanced ECL intensity has a linear relationship with the number of HeLa cells in the range of 5∼5000 and with a detection limit of 2 HeLa cells. The proposed ECL biosensor has high specificity to telomerase in the presence of common interferents. The relative standard deviations (RSDs) were <5 % at 100 HeLa cells. The proposed method provides a convenient approach for telomerase-related cancer screening or diagnosis.
Ministry of Education Key Laboratory of Analysis and Detection for Food Safety, Fujian Provincial Key Laboratory of Analysis and Detection for Food Safety, Fuzhou University, Fuzhou, Fujian, 350116, China.
Yue
Y
Lin
Ministry of Education Key Laboratory of Analysis and Detection for Food Safety, Fujian Provincial Key Laboratory of Analysis and Detection for Food Safety, Fuzhou University, Fuzhou, Fujian, 350116, China.
Linlin
L
Yang
Ministry of Education Key Laboratory of Analysis and Detection for Food Safety, Fujian Provincial Key Laboratory of Analysis and Detection for Food Safety, Fuzhou University, Fuzhou, Fujian, 350116, China.
Guiyin
G
Yue
State Key Laboratory of Structural Chemistry, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fuzhou, Fujian, 350002, China.
Lifen
L
Chen
Ministry of Education Key Laboratory of Analysis and Detection for Food Safety, Fujian Provincial Key Laboratory of Analysis and Detection for Food Safety, Fuzhou University, Fuzhou, Fujian, 350116, China.
Bin
B
Qiu
Ministry of Education Key Laboratory of Analysis and Detection for Food Safety, Fujian Provincial Key Laboratory of Analysis and Detection for Food Safety, Fuzhou University, Fuzhou, Fujian, 350116, China.
Longhua
L
Guo
Ministry of Education Key Laboratory of Analysis and Detection for Food Safety, Fujian Provincial Key Laboratory of Analysis and Detection for Food Safety, Fuzhou University, Fuzhou, Fujian, 350116, China. zylin@fzu.edu.cn.|Department of Chemistry, Fuzhou University, Fuzhou, Fujian, 350116, China. zylin@fzu.edu.cn.
Zhenyu
Z
Lin
Ministry of Education Key Laboratory of Analysis and Detection for Food Safety, Fujian Provincial Key Laboratory of Analysis and Detection for Food Safety, Fuzhou University, Fuzhou, Fujian, 350116, China.
Guonan
G
Chen
D015374:Biosensing Techniques / Q000379:methods; D004247:DNA / Q000378:metabolism*; D057075:Enzyme Assays / Q000379:methods; D006367:HeLa Cells; D006801:Humans; D007364:Intercalating Agents / Q000378:metabolism*; D049408:Luminescent Agents / Q000378:metabolism*; D008163:Luminescent Measurements / Q000379:methods*; D009942:Organometallic Compounds / Q000378:metabolism*; D010618:Phenanthrolines / Q000378:metabolism*; D019098:Telomerase / Q000378:metabolism*
D007364:Intercalating Agents; D049408:Luminescent Agents; D009942:Organometallic Compounds; D010618:Phenanthrolines; C034282:tris-(1,10-phenanthroline)ruthenium; D004247:DNA; D019098:Telomerase
Electrochemiluminescence; HeLa cell; Label free; Telomerase