| Summary: | In this study, magnetic maghemite nanoparticles, which belong to the group of metal oxides, were functionalized with chitosan, a non-toxic, hydrophilic, biocompatible, biodegradable biopolymer with anti-bacterial effects. This was done using different synthesis methods, and a comparison of the properties of the synthesized chitosan functionalized maghemite nanoparticles was conducted. Characterization was performed using scanning electron microscopy (SEM) and vibrating sample magnetometry (VSM). Characterizations of size distribution were performed using dynamic light scattering (DLS) measurements and laser granulometry. A chitosan functionalization layer was confirmed using potentiometric titration on variously synthesized chitosan functionalized maghemite nanoparticles, which is important for further immobilization of bioactive compounds. Furthermore, after activation of chitosan functionalized maghemite nanoparticles with glutaraldehyde (GA) or pentaethylenehexamine (PEHA), immobilization studies of enzyme cholesterol oxidase (ChOx) and horseradish peroxidase (HRP) were conducted. Factors influencing the immobilization of enzymes, such as type and concentration of activating reagent, mass ratio between carrier and enzyme, immobilization time and enzyme concentration, were investigated. Briefly, microparticles made using the chitosan suspension cross-linking process (MC2) proved to be the most suitable for obtaining the highest activity of immobilized enzyme, and nanoparticles functionalized with chitosan using the covalent binding method (MC3) could compete with MC2 for their applications.
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