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- 저자 : T. Jouenne (검색결과 2 건)
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Monitoring of E. coli Immobilization on Modified Gold Electrode: A New Bacteria-based Glucose Sensor
N. Borghol,A. Othmane,L. Mora,T. Jouenne,A. C. Duncan,N. Jaffézic-Renault,N. Sakly,Y. Chevalier,P. Lejeune 한국생물공학회 2010 Biotechnology and Bioprocess Engineering Vol.15 No.2
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 (Ret) 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 Ret 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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N. Zanina,L. Mora,A. Othmane,M. Bénard,A. Duncan,T. Jouenne,D. Vaudry,M. Souiri 한국생물공학회 2013 Biotechnology and Bioprocess Engineering Vol.18 No.1
Metastasis mechanisms depend on cell metabolism changes, migration and adhesion to different tissues. To understand their choice of interaction site, the tumoral cell adhesion to model surfaces was studied. The response of Caco-2 tumoral cells cultured on polyelectrolyte filmfunctionalized surfaces with or without adhesion proteins (fibronectin or collagen IV) was analyzed. Using the layerby-layer method, multilayer films were prepared with cationic poly(allylamine hydrochloride) and anionic poly(sodium 4-styrenesulfonate) polyelectrolytes. Film surface wettability was evaluated. The electrochemical impedance spectroscopy analyses were carried out to control the elaborated surfaces on which Caco-2 tumoral cells were cultured. The cell velocity was studied by video-microscopy and a cell colorimetric assay (WST-1) was used to quantify cell viability. The film surface parameters as well as the protein nature and localization in the film were found to modulate cell response. Results demonstrated that the cancer cell motility and proliferation were higher when cultured onto pure collagen located above the polyelectrolyte film and that the reverse surprisingly was observed when proteins were inserted into the polyelectrolyte film. Data also showed that cell motility was correlated with a high charge transfer resistance (Rct) and a low surface free energy (SFE)polar component (electron donor character). This relationship was valid only for pure external proteins. Thus, fibronectin exhibited a low Rct and a high SFE polar component,which decreased cell motility and proliferation.
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