A Highly Sensitive Electrochemical Immunosensor for the detection of CCR4 Biomarker with Star-Shaped Polymer Covered Disposable Electrode

Abstract

An electrochemical immunosensor prepared with the use of star-shaped polymer (StrPol(GMA)) as a biosensor matrix material was manufactured to quantify the CC chemokine receptor 4 (CCR4) molecules in human serum. Various characterization techniques, including electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV), and scanning electron microscopy (SEM), were used for the analysis of the fabrication steps of the proposed system. The impedance and CV analyses at each modification step of the manufacturing process illustrated the charge transport kinetics of the ferri-ferro redox solution. The binding of anti-CCR4 antibody and the related antibody-its CCR4 antigen biointeraction on the working electrode surface changed the interfacial electron transport. The interactions of anti-CCR4 with different levels of CCR4 were further followed via variations in impedance responses. The results illustrated that the electron transport resistance rose linearly with rising amounts of CCR4 antigen. The dynamic range and detection limit (LOD) were determined as 0.115-23 pg/mL and 34.9 fg/mL, respectively. The specificity, regeneration, storage stability, and reproducibility tests were also performed. Furthermore, CCR4 detection in the presence of interfering biomarkers was accomplished with success using the biosensing approach. The results of these studies illustrate that the introduced biosensor was a useful tool for monitoring low concentrations of CCR4 in human sera.

Keywords

• CC chemokines receptor 4
• prostate cancer
• disposable biosensor

References

1. S. Singh, A.A. Gill, M. Nlooto, R. Karpoormath, Prostate cancer biomarkers detection using nanoparticles based electrochemical biosensors, Biosens. Bioelectron., 137 (2019) 213-221.
2. E.B. Aydın, M. Aydın, M.K. Sezgintürk, Fabrication of electrochemical immunosensor based on acid-substituted poly (pyrrole) polymer modified disposable ITO electrode for sensitive detection of CCR4 cancer biomarker in human serum, Talanta 222 (2021) 121487.
3. J.A. Halpern, C. Oromendia, J.E. Shoag, S. Mittal, M.F. Cosiano, K.V. Ballman, A.J. Vickers, J.C. Hu, Use of digital rectal examination as an adjunct to prostate specific antigen in the detection of clinically significant prostate cancer, J. Urol., 199(4) (2018) 947-953.
4. L. Xu, Y. Wen, S. Pandit, V.R. Mokkapati, I. Mijakovic, Y. Li, M. Ding, S. Ren, W. Li, G. Liu, Graphene-based biosensors for the detection of prostate cancer protein biomarkers: a review, BMC Chem., 13 (2019) 1-12.
5. E.B. Aydın, M. Aydın, M.K. Sezgintürk, Ultrasensitive and Selective Impedimetric Determination of Prostate Specific Membrane Antigen Based on Di‐Succinimide Functionalized Polythiophene Covered Cost‐Effective Indium Tin Oxide, Macromol. Biosci., 21(10) (2021) 2100173.
6. E.B. Aydın, M. Aydın, M.K. Sezgintürk, A highly sensitive immunosensor based on ITO thin films covered by a new semi-conductive conjugated polymer for the determination of TNFα in human saliva and serum samples, Biosens. Bioelectron., 97 (2017) 169-176.
7. E.B. Aydın, M. Aydın, M.K. Sezgintürk, Highly sensitive electrochemical immunosensor based on polythiophene polymer with densely populated carboxyl groups as immobilization matrix for detection of interleukin 1β in human serum and saliva, Sens. Actuators, B, 270 (2018) 18-27.
8. E. Turan, A. Zengin, Z. Suludere, N.Ö. Kalkan, U. Tamer, Construction of a sensitive and selective plasmonic biosensor for prostate specific antigen by combining magnetic molecularly-imprinted polymer and surface-enhanced Raman spectroscopy, Talanta, 237 (2022) 122926.

9. M. Tinzl, M. Marberger, S. Horvath, C. Chypre, DD3PCA3 RNA analysis in urine–a new perspective for detecting prostate cancer, Eur. Urol., 46(2) (2004) 182-187.
10. E.B. Aydın, M. Aydın, M.K. Sezgintürk, Electrochemical immunosensor for detection of CCR4 cancer biomarker in human serum: An alternative strategy for modification of disposable ITO electrode, Macromol. Biosci., 21(2) (2021) 2000267.
11. J. Vandercappellen, J. Van Damme, S. Struyf, The role of CXC chemokines and their receptors in cancer, Cancer lett., 267(2) (2008) 226-244.
12. E.T. da Silva, D.E. Souto, J.T. Barragan, J. de F. Giarola, A.C. de Moraes, L.T. Kubota, Electrochemical biosensors in point‐of‐care devices: recent advances and future trends, ChemElectroChem, 4(4) (2017) 778-794.
13. L. Jing, C. Xie, Q. Li, M. Yang, S. Li, H. Li, F. Xia, Electrochemical biosensors for the analysis of breast cancer biomarkers: From design to application, Anal. Chem., 94(1) (2021) 269-296.
14. F. Teles, L. Fonseca, Applications of polymers for biomolecule immobilization in electrochemical biosensors, Mater. Sci. Eng. C., 28(8) (2008) 1530-1543.
15. S. Shrivastava, N. Jadon, R. Jain, Next-generation polymer nanocomposite-based electrochemical sensors and biosensors: A review, TrAC, Trends Anal. Chem., 82 (2016) 55-67.
16. M. Aydın, E.B. Aydın, M.K. Sezgintürk, A disposable immunosensor using ITO based electrode modified by a star-shaped polymer for analysis of tumor suppressor protein p53 in human serum, Biosens. Bioelectron., 107 (2018) 1-9.
17. J.M. Ren, T.G. McKenzie, Q. Fu, E.H. Wong, J. Xu, Z. An, S. Shanmugam, T.P. Davis, C. Boyer, G.G. Qiao, Star polymers, Chem. Rev., 116(12) (2016) 6743-6836.
18. S. Sánchez, M. Roldán, S. Pérez, E. Fàbregas, Toward a fast, easy, and versatile immobilization of biomolecules into carbon nanotube/polysulfone-based biosensors for the detection of hCG hormone, Anal. Chem., 80(17) (2008) 6508-6514.
19. F. Mollarasouli, S. Kurbanoglu, S.A. Ozkan, The role of electrochemical immunosensors in clinical analysis, Biosensors, 9(3) (2019) 86.
20. E. Emregul, O. Kocabay, B. Derkus, T. Yumak, K.C. Emregul, A. Sınag, K. Polat, A novel carboxymethylcellulose–gelatin–titanium dioxide–superoxide dismutase biosensor; electrochemical properties of carboxymethylcellulose–gelatin–titanium dioxide–superoxide dismutase, Bioelectrochemistry, 90 (2013) 8-17.
21. J.P. Kunhan, P.S. Chandrappa, C. Ravikumar, N. Hanumantharayappa, R. Naik, R. Pothu, R. Boddula, A. Al Otaibi, Study of cobalt doped gdalo3 for electrochemical application, Curr. Anal. Chem., 17(5) (2021) 662-667.
22. M. Aydın, An ultrasensitive immunosensor based on tri-armed star poly (glycidyl methacrylate) polymer-coated ITO-PET electrode for detection of neuron-specific enolase in human serum, Int. J. Environ. Anal. Chem., 100(4) (2020) 492-506.
23. J. Oh, J.-H. Lee, J.C. Koo, H.R. Choi, Y. Lee, T. Kim, N.D. Luong, J.-D. Nam, Graphene oxide porous paper from amine-functionalized poly (glycidyl methacrylate)/graphene oxide core-shell microspheres, J. Mater. Chem., 20(41) (2010) 9200-9204.
24. A. Banas, K. Banas, A. Furgal-Borzych, W.M. Kwiatek, B. Pawlicki, M. Breese, The pituitary gland under infrared light–in search of a representative spectrum for homogeneous regions, Analyst, 140(7) (2015) 2156-2163.
25. Z. Ganim, H.S. Chung, A.W. Smith, L.P. DeFlores, K.C. Jones, A. Tokmakoff, Amide I two-dimensional infrared spectroscopy of proteins, Acc. Chem. Res., 41(3) (2008) 432-441.
26. N.F. Atta, A. Galal, S.M. Ali, D.M. El-Said, Improved host–guest electrochemical sensing of dopamine in the presence of ascorbic and uric acids in a β-cyclodextrin/Nafion®/polymer nanocomposite, Anal. Meth., 6(15) (2014) 5962-5971.
27. J. Eveness, L. Cao, J. Kiely, R. Luxton, Equivalent circuit model of a non-faradaic impedimetric ZnO nano-crystal biosensor, J. Electroanal. Chem., 906 (2022) 116003.
28. T. Bertok, L. Lorencova, E. Chocholova, E. Jane, A. Vikartovska, P. Kasak, J. Tkac, Electrochemical impedance spectroscopy based biosensors: Mechanistic principles, analytical examples and challenges towards commercialization for assays of protein cancer biomarkers, ChemElectroChem, 6(4) (2019) 989-1003.
29. M. Yuksel, W. Luo, B. McCloy, J. Mills, M. Kayaharman, J.T. Yeow, A precise and rapid early pregnancy test: Development of a novel and fully automated electrochemical point-of-care biosensor for human urine samples, Talanta, 254 (2023) 124156.