科研之友
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摘要
This article deals with the elaboration and characterization of an original and easy way to produce templates able to host dyes and enzymes. The template films are made of plasma methacrylic acid (MAA) and ethylene glycol dimethacrylate (EGDMA) copolymers and allow for the design of a free-standing matrix able to host active enzymes. Plasma copolymers based on 80/20% molar ratio MAA and EGDMA were prepared to obtain stable chemical plasma films containing carboxyl groups. The amount of such functional groups was quantified by chemical derivatization, Fourier transform infrared spectroscopy and nuclear magnetic resonance spectroscopies. The analysis reveals the possibility to produce stable coatings with accessible functional groups to control the affinity of the film for dyes such as toluidine blue and enzymes such as alkaline phosphatase. We showed that this kind of plasma film is able to entrap enzymes whose catalytic activity is still preserved. Enzymatic activity was studied by measuring the production of paranitrophenol from the hydrolysis of paranitrophenyl phosphate (PNP). This distribution of the enzyme across the film thickness was investigated by means of laser confocal scanning microscopy and time of flight secondary ion mass spectrometry. Some desorption of enzyme was observed during and after the first enzymatic assay, but stable activity was obtained thereafter. The mechanism by which the negatively charged enzyme is entrapped in the plasma polymer matrix, also carrying negatively charged groups, is briefly discussed.
- 出版日期2012-10-11
