A new self-assembled layer-by-layer glucose biosensor based on chitosan biopolymer entrapped enzyme with nitrogen doped graphene.

The layer-by-layer (LbL) technique has been used for the construction of a new enzyme biosensor. Multilayer films containing glucose oxidase, GOx, and nitrogen-doped graphene (NG) dispersed in the biocompatible positively-charged polymer chitosan (chit(+)(NG+GOx)), together with the negatively charged polymer poly(styrene sulfonate), PSS(-), were assembled by alternately immersing a gold electrode substrate in chit(+)(NG+GOx) and PSS(-) solutions. Gravimetric monitoring during LbL assembly by an electrochemical quartz microbalance enabled investigation of the adsorption mechanism and deposited mass for each monolayer. Cyclic voltammetry and electrochemical impedance spectroscopy were used to characterize the LbL modified electrodes, in order to establish the contribution of each monolayer to the overall electrochemical properties of the biosensor. The importance of NG in the biosensor architecture was evaluated by undertaking a comparative study without NG in the chit layer. The GOx biosensor's analytical properties were evaluated by fixed potential chronoamperometry and compared with similar reported biosensors. The biosensor operates at a low potential of -0.2V vs., Ag/AgCl, exhibiting a high sensitivity of 10.5 µA cm(-2) mM(-1), and a detection limit of 64 µM. This study shows a simple approach in developing new biosensor architectures, combining the advantages of nitrogen-doped graphene with the LbL technique for enzyme immobilization.

Comparative spectroscopic studies on liposomes containing chlorophyll a and chlorophyllide a.

Chlorophyll a (Chla) and chlorophyllyde a (Chlida) - a derivative of Chla, have been incorporated in the lipid bilayers of two types of liposomes, small unilamellar vesicles (SUV) and multilamelar vesicles (MLV). The objective of the present work was to compare the spectral behaviour of Chla and Chlida incorporated in the lipid bilayers and their sensing behaviour at molecular level. The VIS absorption and fluorescence emission presented differences depending on the type of liposomes and inserted pigment, reflecting the different localization of porphyrin macrocycle in the lipid moieties. The temperature dependence of emission anisotropy and fluorescence intensity, for both Chla and Chlida incorporated in DPPC SUV, revealed the presence of different lipid phases. The degree of incorporation of quercetin (QCT) in liposome membrane was studied by using Chla and Chlida as molecular sensors. The fluorescence polarisation data and the fluorescence quenching process provided arguments for the insertion of the QCT at the interface lipid/water, in the vicinity of lipid polar heads and porphyrin macrocycle. The phytyl chain of Chla penetrating in the hydrophobic core of the lipid bilayers is responsible for the observed differences among Chla and Chlida in sensing the lipid phase transition and the fluorescence quenching process induced by QCT.

The effects of the natural antioxidant quercetin and anions of the Hofmeister series on liposomes marked with chlorophyll a.

The unilamellar liposomes of dimyristoylphosphatidylcholine (DMPC), marked with chlorophyll a (Chla), have been chosen as suitable models in studies aimed at determining the effects of natural antioxidant quercetin (QCT) and Hofmeister series anions on lipid bilayers. The variations of steady state fluorescence emission intensity of Chla have been recorded and the values of Chla fluorescence anisotropy, under constant temperature and viscosity, at pH = 7.3-7.4 have been measured. Two types of experiments have been performed. In the first type of experiment, the concentration of anions was maintained constant and the concentration of QCT was varied (from 0 to 100 micromol/l). In the second type of experiment, the concentration of QCT was constant (30 micromol/l) and the concentration of anions was varied (from 0 to 152 mmol/l). The quenching of Chla fluorescence by QCT pleads in favor of QCT insertion at interface water-lipid, in the vicinity of the polar heads of lipids from liposomal bilayer at physiological pH and temperature. Fluorescence anisotropy of Chla in liposomes brought more evidences for QCT localizations at lipid/water interface. Chla is sensing a more rigid microenvironment when QCT is added to the lipid bilayer and specific effects of the Hofmeister series anions.

Interaction of imatinib with liposomes: voltammetric and AFM characterization.

The interaction of imatinib with 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and 2-oleoyl-1-stearoyl-sn-glycero-3-phosphocholine (OSPC) liposomes and the adsorption of DPPC and OSPC were studied using atomic force microscopy (AFM) at highly oriented pyrolytic graphite (HOPG) and differential pulse voltammetry at glassy carbon electrode (GCE). The HOPG induces the rupture of the liposomes and allows the lipids to adsorb along one of the three axes of symmetry of the HOPG basal planes, forming well-ordered lamellar structures. After interaction, both DPPC monolayers and DPPC-imatinib complexes are adsorbed onto HOPG. The OSPC-imatinib complexes self-organize only into ordered but larger domains of parallel stripes that maintain the threefold symmetry of the HOPG, due to an easier imatinib penetration into the unsaturated OSPC liposome bilayers. The voltammetric results show that upon interaction, the electrochemical active moiety of imatinib is incorporated into the lipid bilayer becoming unavailable to the GCE surface for oxidation, leading to local structural modifications of the lipid bilayer which were also electrochemically detected. A model is proposed for the liposome-imatinib interaction considering that imatinib interacts primarily by van der Waals and hydrogen bonds with the phosphatidylcholine headgroups, leading to defects in the liposome bilayer and allowing further incorporation of imatinib into the liposome lamellae.