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IV. Réalisation de membranes enzymatiques épaisses pour microcapteurs ENFETs 70 60 (b) 50 C(buffer)=lmM r° 30 20 C(b u ife r)= 10m M r-D 10 C(buffer)=50 o 5 lO 15 20 25 30 C(BuChCI) (mM) Fig. 3. Response of the butyryicholinesterase biosensor versus buffer concentration. (a)for BuChE-BSA FET: C (bufjèr) =1mM; C (buffer) =5mM; C (buffer) =10mM; (b) for BuChE-PVA/SbQ FET: C (buffer) =1mM; C (buffer) =10mM; AC (buffer) =50mM Figures 2, represents the sensitivity of two different type ENFETs vs. the pH of the 1 mM phosphate buffer. Higher sensitivity is obtained at pH equal to 8-8.5 whatever the membrane used. The same tendency was obtained for acetyicholinesterase based sensors [8, 9], Figures 3a and 3b represent the sensitivity of ENFETs vs. the concentration of the phosphate buffer. lt was found firstly that the response of ENFETs could be changed by varying the concentration of the buffer and secondly that the both enzymatic membranes were more sensitive in 1 mM buffer (lower concentration tested). lt is noteworthy that a lower concentration would not allow to maintain a constant ionic strength during liberation of H, which takes place during the enzymatic reaction. In this context, the pH and the ionic strength selected for further experiments were phosphate buffer of concentration 1mM at pH=8.O. 3.2. Biosensor calibration The biosensor calibration curves were presented following two basic methods : first, the steadystate sensor response was registered (Fig. 4a and 5a) and secondly, the kinetìc response (Fig. 4b and 5b). Fig. 4a and 5a show that the dynamic ranges for both membranes are very similar. In addition, a very low standard deviation is obtained for the both curves (less than 1%). On opposite, the curves 128
IV. Réalisation de membranes enzymatiques épaisses pour microcapteurs ENFETs obtained in the kinetic mode (Fig. 4b and 5b) show that the dynamic linear ranges differ strongly: 0.2-1 mM for BSA membrane and 0.2-5.8 mM for PVAISbQ membrane. For the latter mode, a great standard deviation is obtained (more than 10%). o s 10 15 20 qßiiaicl) (mM) 2500 2000 . 1500 e 1000 500 Km 0 2 4 6 8 lO 12 14 16 8 20 C(BuChCI) (mM) Fig. 4. Calibration curves for BuChE-BSA FET in 1 mM phosphate buffèr pH=8: (a) stationary mode; (b) kinetic mode. 129
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nt y (d) N° d'ordre: ECL 99-28 Ann
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ECOLE CENTRALE DE LYON LISTE DES PE
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ECOLE CENTRALE DE LYON LISTE DES PE
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Sommaire SOMMAiRE INTRODUCTION GENE
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Sommaire REFERENCE BIBLIOGRAPHIQUES
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INTRODUCTION GENERALE
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Introduction générale un capteur
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I. Les biocapteurs Les biocapteurs
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I. Les biocapteurs Le biocapteur is
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I. Les biocapteurs LIGAND Enzymes S
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L Les biocapteurs pour l'obtention
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I. Les biocapteurs Dans l'état act
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I. Les biocapteurs au sein d'une ma
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I. Les biocapteurs 5. Couplage cova
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L Les biocapteurs 1.4.4. La réticu
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I. Les biocapteurs Tableau 1.3. Car
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I. Les bloca pfeurs 2. - Les oxydor
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I. Les biocapteurs 11.3. La cinéti
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I. Les biocapteurs A t état initia
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I. Les biocapteurs atteindre une vi
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I. Les biocapteurs La représentati
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I. Les biocapteurs Dans l'inhibitio
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I. Les biocapteurs 11.5.1. Influenc
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I. Les biocapteurs REFERENCES BIBLI
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CHAPITRE II LE TRANSDUCTEUR ET LA B
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Il. Le transducteur et la biofoncti
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II. Le transducteur et la biofoncti
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Il. Le transducteur et la bio fonct
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e .20 -to 5 I 10 o 20 Ttrne minutes
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CHAPITRE III Etudes du film Langmui
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Ill. Etudes du film Lan gmuir-Blodg
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III. Etudes du film Lan gmuir-Bi'od
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III. Etudes du film Lan gmuir-Blodg
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III. Etudes du film Lan gmuir-Blodg
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Ill. Etudes du film Lan gmuir-Blodg
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