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Immobilisation de la trypsine sur un support de polyéthylène fonctionnalisé par voie plasma

Mahsa Ghasemi 
Abstract : Cold plasma processing is a very efficient technique to modify the surface of polymers without changing their bulk properties. By this way, the immobilization of enzymes on inert supports becomes possible. In this work different plasma processes (mixture of nitrogen and hydrogen, ammonia, and allylamine) were selected in order to generate primary amine functions on the surface of low density polyethylene (PE) films. The activation of the surface by amino groups was followed by a reaction with glutaraldehyde as homobifunctional cross-linking agent leading to the first imine linkage. The enzyme was then reacted with free aldehyde functions and fixed to the surface via lysine residues (imine bond). In some cases, the reduction of the imine bonds by sodium cyanoborohydride was performed to form the stable secondary amines. The activity of the trypsin immobilized on the PE disks was assayed by kinetic measurement of N-α-Benzoyl-L-Arginine Ethyl Ester (BAEE) hydrolysis. The surface composition of the modified polyethylene samples was characterized by X-ray photoelectron spectroscopy (XPS) and confirms the composition of the different layers fixed on to the surface of PE. In order to check the strength of immobilization, different washing solutions were used. Active trypsin was successfully immobilized on the surface of polyethylene with a maximal value of activity equal to 0.080 U/cm2 and no enzyme was released during the reaction. We also studied the stability of the immobilized enzyme during repetitive assays (4 assays) which showed a decrease of activity over a period of two months.
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Submitted on : Monday, May 4, 2009 - 8:00:00 AM
Last modification on : Monday, May 4, 2009 - 8:00:00 AM
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  • HAL Id : pastel-00005078, version 1



Mahsa Ghasemi. Immobilisation de la trypsine sur un support de polyéthylène fonctionnalisé par voie plasma. Engineering Sciences [physics]. Chimie ParisTech, 2007. English. ⟨pastel-00005078⟩



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