Mechanisms of Influenza Virus HA2 Peptide Interaction with Liposomes Studied by Dual-Wavelength MP-SPR.

A phospholipid-based liposome layer was used as an effective biomimetic membrane model to study the binding of the pH-dependent fusogenic peptide (E4-GGYC) from the influenza virus hemagglutinin HA2 subunit. To this end, a multiparameter surface plasmon resonance approach (MP-SPR) was used for monitoring peptide-liposome interactions at two pH values (4.5 and 8) by means of recording sensorgrams in real time without the need for labeling. Biotinylated liposomes were first immobilized as a monolayer onto the surface of an SPR gold chip coated with a streptavidin layer. Multiple sets of sensorgrams with different HA2 peptide concentrations were generated at both pHs. Dual-wavelength Fresnel layer modeling was applied to calculate the thickness (d) and the refractive index (n) of the liposome layer to monitor the change in its optical parameters upon interaction with the peptide. At acidic pH, the peptide, in its α helix form, entered the lipid bilayer of liposomes, inducing vesicle swelling and increasing membrane robustness. Conversely, a contraction of liposomes was observed at pH 8, associated with noninsertion of the peptide in the double layer of phospholipids. The equilibrium dissociation constant KD = 4.7 × 10-7 M of the peptide/liposome interaction at pH 4.5 was determined by fitting the "OneToOne" model to the experimental sensorgrams using Trace Drawer software. Our experimental approach showed that the HA2 peptide at a concentration up to 100 µM produced no disruption of liposomes at pH 4.5.

Exploring the benefits of surface analysis techniques to develop double multilayer transfer printing of J-Aggregates cyanine dyes by integrating L-b-L and µCp processes.

Layer-by-layer self-assembly (L-b-L assembly) makes possible to obtain polyelectrolyte multilayers (PEMs) and one of the polyelectrolytes could be replaced by a dye molecule to obtain multilayers which may exhibit optical properties of great interest. On the other hand, µCp has become a routine technique for the preparation of micro- and nanostructured surfaces. In our development in progress of a surface engineering strategy to transfer J-Agg cyanine dyes onto surfaces by integrating L-b-L process and µCp, this contribution highlights how surface analysis imaging techniques can bring valuable information for the development of the process involving a double Multilayers Transfer Printing (MTP) with a Moiré effect. Key parameters sustaining image interpretation are difference in deposit thickness (optical microscopy, atomic force microscopy, scanning electron microscopy), in roughness (atomic force microscopy and scanning electron microscopy), in charge effect (scanning electron microscopy) and the chemical contrast between unprinted and printed areas (time-of-flight secondary ion mass spectrometry).

Surface Plasmon Resonance Monitoring of Mono-Rhamnolipid Interaction with Phospholipid-Based Liposomes.

The interactions of mono-rhamnolipids (mono-RLs) with model membranes were investigated through a biomimetic approach using phospholipid-based liposomes immobilized on a gold substrate and also by the multiparametric surface plasmon resonance (MP-SPR) technique. Biotinylated liposomes were bound onto an SPR gold chip surface coated with a streptavidin layer. The resulting MP-SPR signal proved the efficient binding of the liposomes. The thickness of the liposome layer calculated by modeling the MP-SPR signal was about 80 nm, which matched the average diameter of the liposomes. The mono-RL binding to the film of the phospholipid liposomes was monitored by SPR and the morphological changes of the liposome layer were assessed by modeling the SPR signal. We demonstrated the capacity of the MP-SPR technique to characterize the different steps of the liposome architecture evolution, i.e., from a monolayer of phospholipid liposomes to a single phospholipid bilayer induced by the interaction with mono-RLs. Further washing treatment with Triton X-100 detergent left a monolayer of phospholipid on the surface. As a possible practical application, our method based on a biomimetic membrane coupled to an SPR measurement proved to be a robust and sensitive analytical tool for the detection of mono-RLs with a limit of detection of 2 µg mL-1.

Voltammetric sensor based on electrodeposited molecularly imprinted chitosan film on BDD electrodes for catechol detection in buffer and in wine samples.

Phenolic compounds such as catechol are present in a wide variety of foods and beverages; they are of great importance due to their antioxidant properties. Their consumption protects against the development of certain diseases such as cancer and cardiovascular diseases. A MIP chitosan (CS) film has been electrodeposited on a boron doped diamond (BDD) electrode, by chronoamperommetry in the presence of catechol, followed by elution with 0.1 M KCl. The morphology of the MIP and non-MIP (NIP) film has been studied by AFM. The electrochemical response of the sensor analyzed by cyclic voltammetry (CV) indicates that the sensor shows excellent reproducibility (RSD = 4.1%) and repeatability (RSD = 7.0%) for catechol detection in the range of 0 to 80 µM, with a detection limit of 6.9 × 10-7 M and high selectivity to catechol recognition versus different phenolic compounds. The results obtained in a red wine show that it can detect catechol in a complex matrix.

Voltammetric Sensor Based on Molecularly Imprinted Chitosan-Carbon Nanotubes Decorated with Gold Nanoparticles Nanocomposite Deposited on Boron-Doped Diamond Electrodes for Catechol Detection.

Phenolic compounds such as catechol are present in a wide variety of foods and beverages; they are of great importance due to their antioxidant properties. This research presents the development of a sensitive and biocompatible molecular imprinted sensor for the electrochemical detection of catechol, based on natural biopolymer-electroactive nanocomposites. Gold nanoparticle (AuNP)-decorated multiwalled carbon nanotubes (MWCNT) have been encapsulated in a polymeric chitosan (CS) matrix. This chitosan nanocomposite has been used to develop a molecular imprinted polymers (MIP) in the presence of catechol on a boron-doped diamond (BDD) electrode. The structure of the decorated MWCNT has been studied by TEM, whereas the characterization of the sensor surface has been imaged by AFM, demonstrating the satisfactory adsorption of the film and the adequate coverage of the decorated carbon nanotubes on the electrode surface. The electrochemical response of the sensor has been analyzed by cyclic voltammetry (CV) where excellent reproducibility and repeatability to catechol detection in the range of 0 to 1 mM has been found, with a detection limit of 3.7 × 10-5 M. Finally, the developed sensor was used to detect catechol in a real wine sample.