Abstract
Electrochemical impedance spectroscopy (EIS) technique has proved to be an effective method for monitoring the immobilization of various bioactive species such as enzymes, DNA, whole cells, and so forth. In this work we describe the development of an electrochemical whole cell based biosensor. Biotinylated fluorescent E. coli are immobilized onto a cysteamine, Sulfo-NHS-LC-biotin, and avidin modified gold electrodes. Immobilized bacteria are clearly observed using confocal microscopy. Electrochemical measurements are based on the charge-transfer kinetics of [Fe (CN)6]3−/4− redox couple. The experimental impedance data were modelised with a computer. SAM assembly and the subsequent immobilization of bacteria on the gold bare electrodes greatly increased the electrontransfer resistance (R et ) and reduced the constant phase element (CPE). It’s interesting to note, the hard immobilization of bacteria on the surface of electrode and do not remove during measurements. The effect of glucose addition was studied in the range of 10−7 μM to 10 μM. The relation between the evolution of R et and D-glucose concentration was found to be linear for values ranging from 10−5 μM to 10−1 μM and reached saturation for higher concentrations. Such biosensor could be applied to a more fundamental study of cell metabolism and drugs effect.
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Sungbo, C. and T. Hagen (2007) Micro hole-based cell chip with impedance spectroscopy. Biosens. Bioelectron. 22: 1764–1768.
Thedinga, E., A. Kob, H. Holst, A. Keuer, S. Drechsler, R. Niendorf, W. Baumann, I. Freund, M. Lehmann, and R. Ehret (2007) Online monitoring of cell metabolism for studying pharmacodynamic effects. Toxicol. Appl. Pharmacol. 220: 33–44.
Junter, G. A. and T. Jouenne (2004) Immobilized viable microbial cells: from the process to the proteome or the cart before the horse. Biotech. Advances 22: 633–658.
Parvanova-Mancheva, T. and V. Beschkov (2009) Microbial denitrification by immobilized bacteria Pseudomonas denitrificans stimulated by constant electric field. Biochem. Eng. J. 44: 208–213.
Corbisier, P., D. Van Der Lelie, B. Borremans, A. Provoost, V. De Lorenzo, N. L. Brown, J. R. Lloyd, J. L. Hobman, E. Csoregi, G. Johansson, and B. Mattiasson (1999) Whole cell-and protein-based biosensors for the detection of bioavailable heavy metals in environmental samples. Analytica. Chimica. Acta. 387: 235–244.
Fernandez Degiorgi, C., R. A. Pizarro, E. E. Smolko, S. Lora, and M. Carenza (2002) Hydrogels for immobilization of bacteria used in the treatment of metal-contaminated wastes. Radiat. Physis. Chem. 63: 109–113.
Hofmann, M. C., D. Ellersiek, F. Kensy, J. Büchs, W. Mokwa, and U. Schnakenberg (2005) Galvanic decoupled sensor for monitoring biomass concentration during fermentation processes. Sens. Actua. B 111–112: 370–375.
Vitalii, S., W. Howard, and V. J. David (1997) SPR Studies of the non-specific adsorption kinetics of human IgG and BSA on gold surfaces modified by self-assembled monolayer (SAMs). J. Colloid Interface Sci. 185: 94–103.
Corbisier, P., E. Thiry, and L. Diels (1996) Bacterial biosensors for the toxicity assessment of solid wastes. Environ. Toxicol. Water Qual. 11: 171–177.
Prime, K. L. and G. M. Whitesides (1993) Adsorption of proteins onto surfaces containing end-attached oligo (ethylene oxide): a model system using self-assembled monolayers. J. Am. Chem. Soc. 115: 10714–10721.
Choi, S. H., J. W. Lee, and S. J. Sim (2005) Enhanced performance of a surface plasmon resonance immunosensor for detecting Ab-GAD antibody based on the modified self-assembled monolayer. Biosens. Bioelectron. 21: 378–383.
Delamarche, E., B. Michel, C. Gerber, D. Anselmetti, H. J. Guentherod, H. Wolf, and H. Ringsdorf (1994) Real-space observation of nanoscale molecular domains in self-assembled monolayers. Langmuir 10: 2869–2871.
Mrksich, M., L. E. Dike, J. Tien, D. E. Ingber, and G. M. Whitesides (1997) Using microcontact printing to pattern the attachment of mammalian cells to self-assembled monolayers of alkanethiolates on transparent films of gold and silver. Exp. Cell. Res. 235: 305–313.
Choi, J. W., K. W. Park, D. B. Lee, W. Lee, and W. H. Lee (2005) Cell immobilization using self-assembled synthetic oligopeptide and its application to biological toxicity detection using surface plasmon resonance. Biosens. Bioelectron. 20: 2300–2305.
Van Hamersveld, E. H., R. G. J. M. van der Lans, and K. C. A. M. Luyben (1997) Quantification of brewers’ yeast flocculation in a stirred tank: Effect of physical parameters on flocculation. Biotechnol. Bioeng. 56: 190–200.
Wilchek, M. and E. A. Bayer (1988) The avidin-biotin complex in bioanalytical applications. Anal. Biochem. 171: 1–32.
Malaquin, L., C. Vieu, M. Genevieve, Y. Tauran, F. Carcenac, M. L. Pourciel, V. Leberre, and E. Trévisiol (2004) Nanoelectrode-based devices for electrical biodetection in liquid solution. Microelec. Eng. 73–74: 887–892.
Jaffrezic-Renault, N. (2001) New trends in biosensors for organophosphorus pesticides. Sensors 1: 60–74.
Gu, H. Y., A. M. Yu, and H. Y. Chen (2001) Direct electron transfer and characterization of hemoglobin immobilized on a Au colloid-cysteamine modified gold electrode. J. Electroananl. Chem. 516: 119–126.
Chaki, N. K. and K. Vijayamohanan (2002) Self-assembled monolayers as a tunable platform for biosensor applications. Biosens. Bioelectron. 17: 1–12.
Tan, J. L., J. Tien, and C. S. Chen (2002) Microcontact printing of proteins on mixed self-assembled monolayers. Langmuir 18: 519–523.
Wink, T., S. J. van Zuilen, A. Bult, and W. P. van Bennekom (1997) Self-assembled monolayers for biosensors. Analyst. 122: 43–50.
Riklin, A. and I. Willner (1995) Glucose and acetylcholine sensing multiplayer enzyme electrodes of controlled enzyme layer thickness. Anal. Chem. 67: 4118–4126.
Wang, C. C., H. Wang, Zh. Y. Wu, G. L. Shen, and R. Q. Yu (2002) A piezoelectric immunoassay based on self assembled monolayers of cystamine and polystyrene sulfonate for determination of schistosoma japonicum antibodies. Anal. Bioanal. Chem. 373: 803–809.
Haugland, R. P. and W. W. You (1995) Coupling of monoclonal antibodies with biotin. Methods Mol. Biol. 45: 223–233.
Spinke, J., M. Liley, F. J. Schmitt, H. J. Guder, L. Angermaier, and W. Knoll (1993) Molecular recognition at self-assembled monolayers: Optimization of surface functionalization. J. Chem. Phys. 99: 7012–7019.
Vidal, O., R. Longin, C. Prigent-Combaret, C. Dorel, M. Hooreman, and P. Lejeune (1998) Isolation of an Escherichia coli K-12 mutant strain able to form biofilms on inert surfaces: involvement of a new ompR allele that increases curli expression. J. Bacteriol. 180: 2442–2449.
Navratilova, I. and P. Skladal (2004) The immunosensors for measurement of 2,4-dichlorophenoxacetic acid based on electrochemical impedance spectroscopy. Bioelectrochemistry 62: 11–18.
Wang, M., L. Wang, G. Wang, X. Ji, Y. Bai, T. Li, S. Gong, and J. Li (2004) Application of impedance spectroscopy for monitoring colloid Au-enhanced antibody immobilization and antibody-antigen reaction. Biosens. Bioelectron. 19: 575–582.
Cui, X., D. Jiang, P. Diao, J. Li, R. Tong, and X. Wang (1999) Assessing the apparent effective thickness of alkanethiol self-assembled monolayers in different concentrations of Fe(CN)6 3−/Fe(CN)6 4− by ac impedance spectroscopy. J. Electroanal. Chem. 470: 9–13.
Darain, F., D. S. Park, and Y. B. Shim (2004) Development of an immunosensor for the detection of vitellogenin using impedance spectroscopy. Biosens. Bioelectron. 19: 1245–1252.
Yang, L., Y. Li, and G. F. Erf (2004) An integrated array microelectrode-based electrophoresis impedance immunosensor for rapid detection of Escherichia coli O157:H7. Anal. Chem. 76: 1107–1113.
Markovich, I. and D. Mandler (2000) The effect of an alkylsilane monolayer on an indium-tin oxide surface on the electrochemistry of hexacyanoferrate. J. Electroanal. Chem. 484: 194–202.
Macdonald J. R. (1992) Impedance spectroscopy. Annals of Biomedical Engineering, 20: 289–305.
Garcia-Belmonte, G., Z. Pomerantz, J. Bisquert, J. P. Lellouche, and A. Zaban (2004) Analysis of ion diffusion and charging in electronically conducting polydicarbazole films by impedance methods. Electrochim. Acta. 49: 3413–3417.
Yang, L., C. Ruan, and Y. Li (2003) Detection of viable Salmonella typhimurium by impedance measurement of electrode capacitance and medium resistance. Biosens. Bioelectron. 19: 495–502.
Munoz-Berbel, X., N. Viguès, J. Mas, A. Toby, A. Jenkins, and Francisco J. Munoz (2007) Impedimetric characterization of the changes produced in the electrode-solution interface by bacterial attachment. Electrochem. Commun. 9: 2654–2660.
Luong, J. H. T., M. Habibi-Rezaei, J. Meghrous, C. **ao, K. B. Male, and A. Kamen (2001) Monitoring motility, spreading, and mortality of adherent insect cells using an impedance sensor. Anal. Chem. 73: 1844–1848.
Savitri, D. and C. K. Mitra (1999) Modeling the surface phenomena in carbon paste electrodes by low frequency impedance and double-layer capacitance measurements. Bioelectrochem. Bioenerg. 48: 163–169.
Ruan, C. M., L. J. Yang, and Y. B. Li (2002) Immunobiosensor chips for detection of Escherichia coli O157:H7 using electrochemical impedance spectroscopy. Anal. Chem. 74: 4814–4820.
Tlili, C., K. Reybier, A. Géloën, L. Ponsonnet, C. Martelet, H. Ben Ouada, M. Lagarde, and N. Jaffrezic-Renault (2003) Fibroblast cells: A sensing bioelement for glucose detection by impedance spectroscopy. Anal. Chem. 75: 3008–3012.
Kong, T., Y. Chen, Y. Ye, K. Zhang, Z. Wang, and X. Wang (2009) An amperometric glucose biosensor based on the immobilization of glucose oxidase on the ZnO nanotubes. Sen. Actuat. B: Chem. 138: 344–350.
Santhosh, P., K. M. Manesh, S. Uthayakumar, S. Komathi, A. I. Gopalan, and K. P. Lee (2009) Fabrication of enzymatic glucose biosensor based on palladium nanoparticles dispersed onto poly(3,4-ethylenedioxythiophene) nanofibers. Bioelectrochemistry 75: 61–66.
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Borghol, N., Mora, L., Jouenne, T. et al. Monitoring of E. coli immobilization on modified gold electrode: A new bacteria-based glucose sensor. Biotechnol Bioproc E 15, 220–228 (2010). https://doi.org/10.1007/s12257-009-0146-4
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DOI: https://doi.org/10.1007/s12257-009-0146-4