A Novel Superoxide Anion Biosensor for Monitoring Reactive Species of Oxygen Released by Cancer Cells

In this study, novel sensitive and selective hydrogel microstructures to detect superoxide anions released by cancer cells, based on electrochemical biosensors, are proposed. Ferrocene was coupled with superoxide dismutase within a poly(ethylene glycol) diacrylate hydrogel matrix. The pre-polymer solution was patterned by photolithography in gold microelectrodes fabricated on top of glass slides. The biosensor was characterized by electrochemical impedance spectroscopy and cyclic voltammetry, and was able to detect superoxide anions in a wide linear range from 5 to 100 μM, with a low detection limit of 0.001 μM and sensitivity of 14.1 nA μM/mm2. Moreover, the biosensor was able to directly detect reactive oxygen species released from prostate cells. Furthermore, the reproducibility, stability and selectivity of the biosensor achieved better results when compared with the previous report, so this methodology can be used in physiological and pathological detection of reactive oxygen species, providing a powerful platform for clinical diagnostics in the future.

[1]  S. Mitra,et al.  Oxidative stress: an essential factor in the pathogenesis of gastrointestinal mucosal diseases. , 2014, Physiological reviews.

[2]  Min Han,et al.  A superoxide anion biosensor based on direct electron transfer of superoxide dismutase on sodium alginate sol-gel film and its application to monitoring of living cells. , 2012, Analytica chimica acta.

[3]  Jiaqiang Xu,et al.  Reagentless amperometric glucose biosensor based on the immobilization of glucose oxidase on a ferrocene@NaY zeolite composite , 2011 .

[4]  B. C. Green,et al.  ELECTRON PARAMAGNETIC RESONANCE STUDIES ON THE PRODUCTION OF FREE RADICALS IN HYDROGEN PEROXIDE AT LIQUID NITROGEN TEMPERATURE1 , 1961 .

[5]  Peng Huang,et al.  Superoxide dismutase as a target for the selective killing of cancer cells , 2000, Nature.

[6]  Shu Liu,et al.  Ultrahigh performance liquid chromatography-triple quadrupole mass spectrometry inhibitors fishing assay: a novel method for simultaneously screening of xanthine oxidase inhibitor and superoxide anion scavenger in a single analysis. , 2012, Analytica chimica acta.

[7]  W. Pfeil,et al.  Denaturation and renaturation of cytochrome c immobilized on gold electrodes in DMSO-containing buffers , 2004 .

[8]  R. Dean,et al.  Biochemistry and pathology of radical-mediated protein oxidation. , 1997, The Biochemical journal.

[9]  C. Noorden,et al.  Oxygen insensitivity of the histochemical assay of glucose-6-phosphate dehydrogenase activity for the discrimination between nonmalignant and malignant cells. , 1999 .

[10]  B. Halliwell,et al.  Oxygen toxicity, oxygen radicals, transition metals and disease. , 1984, The Biochemical journal.

[11]  Barry Halliwell,et al.  Oxidative stress and neurodegeneration: where are we now? , 2006, Journal of neurochemistry.

[12]  T. Ohsaka,et al.  A carbon fiber microelectrode-based third-generation biosensor for superoxide anion. , 2005, Biosensors & bioelectronics.

[13]  Lu Wang,et al.  A novel amperometric biosensor for superoxide anion based on superoxide dismutase immobilized on gold nanoparticle-chitosan-ionic liquid biocomposite film. , 2013, Analytica chimica acta.

[14]  J. Di,et al.  Third-generation superoxide anion sensor based on superoxide dismutase directly immobilized by sol-gel thin film on gold electrode. , 2004, Biosensors & bioelectronics.

[15]  Adam Heller,et al.  Electrical communication between redox centers of glucose oxidase and electrodes via electrostatically and covalently bound redox polymers , 1989 .

[16]  S. Ray,et al.  Centchroman: A safe reversible postcoital contraceptive with curative and prophylactic activity in many disorders. , 2018, Frontiers in bioscience.

[17]  J. Kehrer.  The Haber-Weiss reaction and mechanisms of toxicity. , 2000, Toxicology.

[18]  F. Rusnak,et al.  What is the ultimate fate of superoxide anion in vivo? , 2002, JBIC Journal of Biological Inorganic Chemistry.

[19]  Alexander Revzin,et al.  Micropatterned Nanocomposite Hydrogels for Biosensing Applications , 2011 .

[20]  C. Malitesta,et al.  Glucose fast-response amperometric sensor based on glucose oxidase immobilized in an electropolymerized poly(o-phenylenediamine) film. , 1990, Analytical chemistry.

[21]  I. Willner,et al.  Probing Biomolecular Interactions at Conductive and Semiconductive Surfaces by Impedance Spectroscopy: Routes to Impedimetric Immunosensors, DNA‐Sensors, and Enzyme Biosensors , 2003 .

[22]  K. B. Oldham,et al.  Use of the ferrocene oxidation process to provide both reference electrode potential calibration and a simple measurement (via semiintegration) of the uncompensated resistance in cyclic voltammetric studies in high-resistance organic solvents , 2000, Analytical chemistry.

[23]  Alexander Revzin,et al.  Immobilizing enzymes onto electrode arrays by hydrogel photolithography to fabricate multi-analyte electrochemical biosensors. , 2010, ACS applied materials & interfaces.

[24]  P. Storz,et al.  Reactive oxygen species in cancer , 2010, Free radical research.

[25]  G. Hotamisligil,et al.  Nutrient sensing and inflammation in metabolic diseases , 2008, Nature Reviews Immunology.

[26]  R. Hudson,et al.  Mass Spectrometry of Inorganic Free Radicals , 1962 .

[27]  T. Tenno,et al.  Superoxide electrode based on covalently immobilized cytochrome c: modelling studies. , 1998, Free radical biology & medicine.

[28]  A. Revzin,et al.  Glucose and lactate biosensors based on redox polymer/oxidoreductase nanocomposite thin films. , 2000, Analytical chemistry.

[29]  C. McNeil,et al.  Sensor-based measurements of the role and interactions of free radicals in cellular systems. , 2002, Journal of biotechnology.

[30]  Xin Wei,et al.  Integration of enzymes and electrodes: spectroscopic and electrochemical studies of chitosan-enzyme films. , 2002, Analytical chemistry.

[31]  L. Andrews.  Infrared spectra and bonding in the sodium superoxide and sodium peroxide molecules , 1969 .

[32]  Pengcheng Xu,et al.  PtW/MoS2 hybrid nanocomposite for electrochemical sensing of H2O2 released from living cells. , 2016, Biosensors & bioelectronics.

[33]  P. Storz.  Reactive oxygen species in tumor progression. , 2005, Frontiers in bioscience : a journal and virtual library.

[34]  R. Haugland,et al.  A stable nonfluorescent derivative of resorufin for the fluorometric determination of trace hydrogen peroxide: applications in detecting the activity of phagocyte NADPH oxidase and other oxidases. , 1997, Analytical biochemistry.

[35]  T. Ohsaka,et al.  Superoxide dismutase-based third-generation biosensor for superoxide anion. , 2002, Analytical chemistry.

[36]  F. Mechta-Grigoriou,et al.  The role of reactive oxygen species and metabolism on cancer cells and their microenvironment. , 2014, Seminars in cancer biology.

[37]  R. Bray,et al.  Oxygen-17 hyperfine splitting in the electron paramagnetic resonance spectrum of enzymically generated superoxide. , 1970, European journal of biochemistry.

[38]  Yang Tian,et al.  Pyramidal, rodlike, spherical gold nanostructures for direct electron transfer of copper, zinc-superoxide dismutase: application to superoxide anion biosensors. , 2008, Langmuir : the ACS journal of surfaces and colloids.

[39]  V. Yadavalli,et al.  Fabrication of poly(ethylene glycol) hydrogel microstructures using photolithography. , 2001, Langmuir : the ACS journal of surfaces and colloids.