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Microchimica Acta

, 186:720 | Cite as

Electrochemical aptamer-based determination of protein tyrosine kinase-7 using toehold-mediated strand displacement amplification on gold nanoparticles and graphene oxide

  • Zongbing Li
  • Zhengkun Zhou
  • Ning Xue
  • Shujie Wu
  • Xiangmin MiaoEmail author
Original Paper

Abstract

An electrochemical method is described for ultrasensitive determination of protein tyrosine kinase-7 (PTK7). It is based on (a) the use of positively charged gold nanoparticles (AuNPs) and negatively charged graphene oxide (GO), and (b) of toehold-mediated strand displacement amplification. A hairpin probe 2 (HP2) containing the sgc8 aptamer was used to modify a glassy carbon electrode (GCE). Its hairpin structure is opened in the presence of PTK7 to form the PTK7-HP2 complex. The exposed part of HP2 partly hybridizes with hairpin probe 1 (HP1) that was immobilizing on the AuNPs and GO modified GCE. On addition of the hairpin probe 3 that was labeled with the redox probe Methylene Blue (MB-HP3), toehold-mediated strand displacement occurs due to complementary hybridization of HP1 with MB-HP3. This causes the release of PTK7-HP2 into the solution and makes it available for the next reaction. Under optimal conditions, PTK7 can be quantified by voltammetry (typically performed at −0.18 V) with a detection limit of 1.8 fM. The assay possesses high selectivity for PTK7 due to the employment of the aptamer. It was successfully applied to the determination of PTK7 in the debris of malignant melanoma A375 cells.

Graphical abstract

Schematic representation of the enzyme-free electrochemical sensor for ultrasensitive determination of protein tyrosine kinase-7 (PTK7) based on the toehold-mediated strand displacement reaction amplification on gold nanoparticles and graphene oxide.

Keywords

Electrochemical sensor Protein tyrosine kinase-7 Positively charged gold nanoparticles Negatively charged graphene oxide Hairpin DNA probes 

Notes

Acknowledgements

This work was supported by the Natural Science Foundation of Xuzhou City (KC18140), and the project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).

Compliance with ethical standards

Conflict of interest

There is no conflict of interest about this article.

Supplementary material

604_2019_3849_MOESM1_ESM.docx (537 kb)
ESM 1 (DOCX 537 kb)

References

  1. 1.
    Saha S, Bardelli A, Buckhaults P, Velculescu VE, Rago C, Croix BS (2001) A phosphatase associated with metastasis of colorectal cancer. Science 294:1343–1346CrossRefGoogle Scholar
  2. 2.
    Kampen KR (2011) Membrane proteins: the key players of a cancer cell. J Membr Biol 242:69–74CrossRefGoogle Scholar
  3. 3.
    Arinaminpathy Y, Khurana E, Engelman DM, Gerstein MB (2009) Computational analysis of membrane proteins: the largest class of drug targets. Drug Discov Today 14:1130–1135CrossRefGoogle Scholar
  4. 4.
    Grimm D, Bauer J, Pietsch J, Infanger M, Eucker J, Eilles C, Schoenberger J (2011) Diagnostic and therapeutic use of membrane proteins in cancer cells. Curr Med Chem 18:176–190CrossRefGoogle Scholar
  5. 5.
    Mothes W, Heinrich SU, Graf R, Nilsson I, Von HG, Brunner J, Rapoport TA (1997) Molecular mechanism of membrane protein integration into the endoplasmic reticulum. Cell 89:523–533CrossRefGoogle Scholar
  6. 6.
    Brown DM, Ruoslahti E (2004) A cell surface protein in breast tumors that mediates lung metastasis. Cancer Cell 5:365–374CrossRefGoogle Scholar
  7. 7.
    Hu MY, Li ZZ, Guo CP, Wang MH, He LH, Zhang ZH (2019) Hollow core-shell nanostructured MnO2/Fe2O3 embedded within amorphous carbon nanocomposite as sensitive bioplatform for detecting protein tyrosine kinase-7. Appl Surf Sci 489:13–24CrossRefGoogle Scholar
  8. 8.
    Wagner G, Peradziryi H, Wehner P, Borchers A (2010) Plexin A1 interacts with PTK7 and is required for neural crest migration. Biochem Biophys Res Commun 402:402–407CrossRefGoogle Scholar
  9. 9.
    Ellington AD, Szostak JW (1990) In vitro selection of RNA molecules that bind specific ligands. Nature 346:818–822CrossRefGoogle Scholar
  10. 10.
    Tuerk C, Gold L (1990) Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase. Science 249:505–510CrossRefGoogle Scholar
  11. 11.
    Shangguan DH, Li Y, Tang ZW, Cao ZHC, Chen HW, Mallikaratchy P, Sefah K, Yang CYJ, Tan WH (2006) Aptamers evolved from live cells as effective molecular probes for cancer study. Proc Natl Acad Sci U S A 103:11838–11843CrossRefGoogle Scholar
  12. 12.
    Li L, Wang Q, Feng J, Tong LL, Tang B (2014) Highly sensitive and homogeneous detection of membrane protein on a single living cell by aptamer and nicking enzyme assisted signal amplification based on microfluidic droplets. Anal Chem 86:5101–5107CrossRefGoogle Scholar
  13. 13.
    Lin S, Gao W, Tian Z, Yang C, Lu L, Mergny JL, Leung CH, Ma DL (2015) Luminescence switch-on detection of protein tyrosine kinase-7 using a G-quadruplex selective probe. Chem Sci 6:4284–4290CrossRefGoogle Scholar
  14. 14.
    Yin JJ, He XX, Wang KM, Qing ZH, Wu X, Shi H, Yang XH (2011) One-step engineering of silver nanoclusters-aptamer assemblies as luminescent labels to target tumor cells. Nanoscale 4:110–112CrossRefGoogle Scholar
  15. 15.
    Miao XM, Li ZB, Zhu AH, Feng ZZ, Tian J, Peng X (2016) Ultrasensitive electrochemical detection of protein tyrosine kinase-7 by gold nanoparticles and methylene blue assisted signal amplification. Biosens Bioelectron 83:39–44CrossRefGoogle Scholar
  16. 16.
    Zheng J, Li NX, Li CR, Wang XX, Liu YC, Mao GB, Ji XH, He ZK (2018) A nonenzymatic DNA nanomachine for biomolecular detection by target recycling of hairpin DNA cascade amplification. Biosens Bioelectron 107:40–46CrossRefGoogle Scholar
  17. 17.
    Li YR, Chang YY, Yuan R, Chai YQ (2018) Highly efficient target recycling-based netlike Y-DNA for regulation of electrocatalysis towards methylene blue for sensitive DNA detection. ACS Appl Mater Interfaces 10:25213–25218CrossRefGoogle Scholar
  18. 18.
    Chen HG, Ren W, Jia J, Feng J, Gao ZF, Li NB, Luo HQ (2016) Fluorometric detection of mutant DNA oligonucleotide based on toehold strand displacement-driving target recycling strategy and exonuclease III-assisted suppression. Biosens Bioelectron 77:40–45CrossRefGoogle Scholar
  19. 19.
    Ding W, Deng W, Zhu H, Liang HJ (2013) Metallo-toeholds: controlling DNA strand displacement driven by Hg(II) ions. Chem Commun 49:9953–9955CrossRefGoogle Scholar
  20. 20.
    Miao XM, Cheng ZY, Ma HY, Li ZB, Xue N, Wang P (2018) Label-free platform for microRNA detection based on the fluorescence quenching of positively charged gold nanoparticles to silver nanoclusters. Anal Chem 90:1098–1103CrossRefGoogle Scholar
  21. 21.
    Qu LL, Wang N, Xu H, Wang WP, Liu Y, Kuo LD, Yadav TP, Wu JJ, Joyner J, Song YH, Li HT, Lou J, Vajtai R, Ajayan PM (2017) Gold nanoparticles and g-C3N4-intercalated graphene oxide membrane for recyclable surface enhanced raman scattering. Adv Funct Mater 27:1701714CrossRefGoogle Scholar
  22. 22.
    Easty DJ, Mitchell PJ, Patel K, Florenes VA, Spritz RA, Bennett DC (1997) Loss of expression of receptor tyrosine kinase family genes PTK7 and SEK in metastatic melanoma. Int J Cancer 71:1061–1065CrossRefGoogle Scholar

Copyright information

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

Authors and Affiliations

  • Zongbing Li
    • 1
  • Zhengkun Zhou
    • 1
  • Ning Xue
    • 1
  • Shujie Wu
    • 1
  • Xiangmin Miao
    • 1
    Email author
  1. 1.School of Life ScienceJiangsu Normal UniversityXuzhouChina

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