Microchimica Acta

, 186:785 | Cite as

A turn-on graphene quantum dot and graphene oxide based fluorometric aptasensor for the determination of telomerase activity

  • Elahe Kazemi
  • Habib BagheriEmail author
  • Dariush Norouzian
Original Paper


A turn-on fluorometric assay is described for determination of the activity of enzyme telomerase. For this purpose, graphene quantum dots (GQDs) were first modified with the telomeric sequence (5′-amino-AATCCGTCGAGCAGAGTT-3′) via a condensation reaction. Injection of graphene oxide causes instant quenching of the blue fluorescence of the GQDs. Addition of cell extract containing telomerase, triggers the extension of telomer via addition of specific sequence (TTAGGG)n to its 3′ end. Fluorescence, best measured at excitation/emission wavelengths of 390/446 nm, is subsequently restored due to folding of the extended telomeric sequence into G-quadruplex structure. The method was applied to the determination of telomerase activity in crude cell extracts of as little as 10 HeLa cells. The linear dynamic range extends from 10 to 6500 cells.

Graphical abstract

In this study, a new turn-on graphene quantum dotm and graphene oxide based fluorometric assay is developed for the determination of telomerase activity


Optical sensing Fluorescence Biosensors Nanoprobe Biomarker Cancer detection G-quadruplex 



This work was supported by the grant number of 95008282 from the Iran National Science Foundation (INSF). The Research Council and Graduates School of Sharif University of Technology (SUT) are also thanked for supporting the project.

Compliance with ethical standards

Competing interests

The author(s) declare that they have no competing interests.

Supplementary material

604_2019_3956_MOESM1_ESM.docx (178 kb)
ESM 1 (DOCX 177 kb)


  1. 1.
    de Lange T (2009) How telomeres solve the end-protection problem. Science 326(5955):948–952. CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Cesare AJ, Reddel RR (2010) Alternative lengthening of telomeres: models, mechanisms and implications. Nat Rev Genet 11:319–330. CrossRefPubMedGoogle Scholar
  3. 3.
    Wang J, Wu L, Ren J, Qu X (2012) Visualizing human telomerase activity with primer-modified Au nanoparticles. Small 8(2):259–264. CrossRefPubMedGoogle Scholar
  4. 4.
    Xiao Y, Dane KY, Uzawa T, Csordas A, Qian J, Soh HT, Daugherty PS, Lagally ET, Heeger AJ, Plaxco KW (2010) Detection of telomerase activity in high concentration of cell lysates using primer-modified gold nanoparticles. J Am Chem Soc 132(43):15299–15307. CrossRefPubMedGoogle Scholar
  5. 5.
    Zhang X, Cheng R, Shi Z, Jin Y (2016) A PCR-free fluorescence strategy for detecting telomerase activity via double amplification strategy. Biosens Bioelectron 75:101–107. CrossRefPubMedGoogle Scholar
  6. 6.
    Zong S, Wang Z, Chen H, Cui Y (2013) Ultrasensitive telomerase activity detection by telomeric elongation controlled surface enhanced raman scattering. Small 9(24):4215–4220. CrossRefPubMedGoogle Scholar
  7. 7.
    Wu L, Wang J, Feng L, Ren J, Wei W, Qu X (2012) Label-free ultrasensitive detection of human telomerase activity using porphyrin-functionalized graphene and electrochemiluminescence technique. Adv Mater 24(18):2447–2452. CrossRefPubMedGoogle Scholar
  8. 8.
    Xiong C, Liang W, Zheng Y, Zhuo Y, Chai Y, Yuan R (2017) Ultrasensitive assay for telomerase activity via self-enhanced electrochemiluminescent ruthenium complex doped metal–organic frameworks with high emission efficiency. Anal Chem 89(5):3222–3227. CrossRefPubMedGoogle Scholar
  9. 9.
    Wang WJ, Li JJ, Rui K, Gai PP, Zhang JR, Zhu JJ (2015) Sensitive electrochemical detection of telomerase activity using spherical nucleic acids gold nanoparticles triggered mimic-hybridization chain reaction enzyme-free dual signal amplification. Anal Chem 87(5):3019–3026. CrossRefPubMedGoogle Scholar
  10. 10.
    Wang F, Gu Z, Lei W, Wang W, Xia X, Hao Q (2014) Graphene quantum dots as a fluorescent sensing platform for highly efficient detection of copper(II) ions. Sensors Actuators B Chem 190:516–522. CrossRefGoogle Scholar
  11. 11.
    Liu X, Wang F, Aizen R, Yehezkeli O, Willner I (2013) Graphene oxide/nucleic-acid-stabilized silver nanoclusters: functional hybrid materials for optical aptamer sensing and multiplexed analysis of pathogenic DNAs. J Am Chem Soc 135(32):11832–11839. CrossRefPubMedGoogle Scholar
  12. 12.
    Morales-Narváez E, Pérez-López B, Pires LB, Merkoçi A (2012) Simple Förster resonance energy transfer evidence for the ultrahigh quantum dot quenching efficiency by graphene oxide compared to other carbon structures. Carbon 50(8):2987–2993. CrossRefGoogle Scholar
  13. 13.
    Qian ZS, Shan XY, Chai LJ, Chen JR, Feng H (2015) A fluorescent nanosensor based on graphene quantum dots–aptamer probe and graphene oxide platform for detection of lead (II) ion. Biosens Bioelectron 68:225–231. CrossRefPubMedGoogle Scholar
  14. 14.
    Loh KP, Bao Q, Eda G, Chhowalla M (2010) Graphene oxide as a chemically tunable platform for optical applications. Nat Chem 2:1015–1024. CrossRefPubMedGoogle Scholar
  15. 15.
    Qian ZS, Shan XY, Chai LJ, Ma JJ, Chen JR, Feng H (2014) A universal fluorescence sensing strategy based on biocompatible graphene quantum dots and graphene oxide for the detection of DNA. Nanoscale 6(11):5671–5674. CrossRefPubMedGoogle Scholar
  16. 16.
    He Y, Zhang B, Fan Z (2018) Aptamer based fluorometric sulfamethazine assay based on the use of graphene oxide quantum dots. Microchim Acta 185(3):163–171. CrossRefGoogle Scholar
  17. 17.
    Mohanty J, Barooah N, Dhamodharan V, Harikrishna S, Pradeepkumar PI, Bhasikuttan AC (2013) Thioflavin T as an efficient inducer and selective fluorescent sensor for the human telomeric G-quadruplex DNA. J Am Chem Soc 135(1):367–376. CrossRefPubMedGoogle Scholar
  18. 18.
    Kazemi E, Dadfarnia S, Haji Shabani AM (2015) Dispersive solid phase microextraction with magnetic graphene oxide as the sorbent for separation and preconcentration of ultra-trace amounts of gold ions. Talanta 141:273–278. CrossRefPubMedGoogle Scholar
  19. 19.
    Liu Z, Robinson JT, Sun X, Dai H (2008) PEGylated nanographene oxide for delivery of water-insoluble cancer drugs. J Am Chem Soc 130(33):10876–10877. CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Ju J, Chen W (2014) Synthesis of highly fluorescent nitrogen-doped graphene quantum dots for sensitive, label-free detection of Fe (III) in aqueous media. Biosens Bioelectron 58:219–225. CrossRefPubMedGoogle Scholar
  21. 21.
    Chang H, Tang L, Wang Y, Jiang J, Li J (2010) Graphene fluorescence resonance energy transfer aptasensor for the thrombin detection. Anal Chem 82(6):2341–2346. CrossRefPubMedGoogle Scholar
  22. 22.
    Zhang M, Yin BC, Tan W, Ye BC (2011) A versatile graphene-based fluorescence “on/off” switch for multiplex detection of various targets. Biosens Bioelectron 26(7):3260–3265. CrossRefPubMedGoogle Scholar
  23. 23.
    Xue Y, Kan ZY, Wang Q, Yao Y, Liu J, Hao YH, Tan Z (2007) Human telomeric DNA forms parallel-stranded intramolecular G-Quadruplex in K+ solution under molecular crowding condition. J Am Chem Soc 129(36):11185–11191. CrossRefPubMedGoogle Scholar
  24. 24.
    Wang L, Chen C, Huang H, Huang D, Luo F, Qiu B, Guo L, Lin Z, Yang H (2018) Sensitive detection of telomerase activity in cancer cells using portable pH meter as readout. Biosens Bioelectron 121:153–158. CrossRefPubMedGoogle Scholar
  25. 25.
    Ou X, Zhan S, Sun C, Cheng Y, Wang X, Liu B, Zhai T, Lou X, Xia F (2019) Simultaneous detection of telomerase and miRNA with graphene oxide-based fluorescent aptasensor in living cells and tissue samples. Biosens Bioelectron 124-125:199–204. CrossRefPubMedGoogle Scholar
  26. 26.
    Wang D, Guo R, Wei Y, Zhang Y, Zhao X, Xu Z (2018) Sensitive multicolor visual detection of telomerase activity based on catalytic hairpin assembly and etching of Au nanorods. Biosens Bioelectron 122:247–253. CrossRefPubMedGoogle Scholar
  27. 27.
    Ding C, Li X, Wang W, Chen Y (2016) Fluorescence detection of telomerase activity in cancer cell extracts based on autonomous exonuclease III-assisted isothermal cycling signal amplification. Biosens Bioelectron 83:102–105. CrossRefPubMedGoogle Scholar
  28. 28.
    He C, Liu Z, Wu Q, Zhao J, Liu R, Liu B, Zhao T (2018) Ratiometric fluorescent biosensor for visual discrimination of cancer cells with different telomerase expression levels. ACS Sens 3(4):757–762. CrossRefPubMedGoogle Scholar
  29. 29.
    Zhang H, Lei Z, Tian R, Wang Z (2018) Polyamidoamine starburst dendrimer-activated chromatography paper-based assay for sensitive detection of telomerase activity. Talanta 178:116–121. CrossRefPubMedGoogle Scholar
  30. 30.
    Zhuang Y, Zhang M, Chen B, Duan R, Min X, Zhang Z, Zheng F, Liang H, Zhao Z, Lou X, Xia F (2015) Quencher group induced high specificity detection of telomerase in clear and bloody urines by AIEgens. Anal Chem 87(18):9487–9493. CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Environmental and Bio-Analytical Laboratories, Department of ChemistrySharif University of TechnologyTehranIran
  2. 2.Pilot Nanobiotechnology DepartmentPasteur Institute of IranTehranIran

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