Synthesis and analytical properties of micrometric biosensing lipobeads

This paper describes the preparation for the first time of lipobead-based micrometric fluorescence biosensors and the optimization of their analytical properties. The study focused on the well-established urea biosensors as a model system. Fluorescence-sensing lipobeads were prepared by coating carboxyl-functionalized silica microspheres with phospholipids. The enzyme urease and the pH indicator fluorescein-5-thiosemicarbazide were then attached covalently to the phospholipid membrane of the lipobeads. Urease converts urea to ammonia, which results in a pH increase in the analyte solution and to a urea concentration-dependent increase in the fluorescence intensity of the sensing lipobeads. Previous fluorescence-sensing lipobeads were synthesized by coating polystyrene particles with a phospholipid membrane. The membrane was physically attached to the particles and the fluorophores were entrapped in the membrane. In this study, we prepared improved fluorescence-sensing lipobeads by utilizing covalent chemistry to bind the phospholipid membrane to the silica particles and the fluorophores to the membrane. This led to improvement in the stability of the newly developed urea-sensing lipobeads compared to previously developed miniaturized fluorescence biosensors.