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Biomedical subjects

C Padeste

Publications and source records attributed to C Padeste.

5 recordsLinked to original sources

Site-directed molecular assembly on templates structured with electron-beam lithography.

We describe a simple method for patterning biomolecular films on surfaces with high resolution. A conventional polymeric resist is structured by electron-beam lithography. The exposed and developed patterns are then used for the directed self-assembly (SA) of a first molecule from solution. Removal of the remaining resist allows the SA of a second species. We illustrate the potential of the approach by assembling on gold (Au) substrates two alkanethiols of contrasting terminal functionality. The patterns have dimensions from the micrometer range down to 40 nm and an edge resolution of 3.5 nm.

Letter↗

Ferrocene-avidin conjugates for bioelectrochemical applications.

Coupling of ferrocene moieties to avidin via a flexible spacer molecule yields a conjugate which combines the unique biotin-binding properties of avidin with the reversible redox characteristics of ferrocenes. Synthesis of the conjugate has been optimised and the conjugates were characterised bio- and electrochemically. Covalent immobilisation of the conjugate on gold electrodes in a dense monolayer results in electrodes with a high binding capacity for biotinylated molecules as well as good electron transfer properties. The application potential of such electrodes for bioelectrochemical systems is demonstrated by electrochemical reduction of hydrogen peroxide under mild conditions catalysed by a bound biotin-microperoxidase MP11 conjugate.

Avidin↗

Neurite outgrowth on microstructured surfaces functionalized by a neural adhesion protein.

Designed networks of neurons are potentially very useful to investigate neural activities. Using photolithography microgrooves suited in size for single neurons have been produced on glass chips. Two conducting gold lanes ending in each microgroove allow extracelluar stimulation of the neurons and recording of their activity. A cell adhesive surface was created by functionalization of glass with the adhesion peptide RGDC. In addition, in order to optimize the contact of the neuronal cell membrane to the electrode surface axonin-1, a specific neural adhesion protein was used. A recombinant form of axonin-1 was produced and immobilized in a correct orientation on protected gold surfaces through a C-terminal cysteine residue. Neurite outgrowth of neurons cultured on chips derivatized with RGDC or axonin-1 were compared. The developed materials and methods represent a first step towards establishing designed functionalized glass surfaces for neurophysiological investigations.

Journal Article↗

Towards amperometric immunosensor devices.

In contrast to optical immunosensors, the electrochemical detection of an immunanalytical reaction does require a labeling, but allows an easier discrimination of specific and non-specific binding. We present a concept and first results for a multivalent amperometric immunosensor system which is based on silicon technology. The capture molecule streptavidin, covalently immobilized on silica, allows the immobilization of biotinylated antigens at a defined density. A nanostructured gold electrode serving as a stable network of nanowires is expected to be beneficial for the electrochemical detection of bound ferrocene-labeled antibody molecules. The results presented focus on site-specific immobilization of streptavidin on silica and reduction of non-specific binding of proteins.

Bacterial Proteins↗

Immobilization of streptavidin for immunosensors on nanostructured surfaces.

A modular immunosensor device based on silicon technology with electrochemical detection is presented. The sensor surface is functionalized with the antigen estradiol using the streptavidin-biotin system. The immobilized antigen is recognized by ferrocene-labeled antibody molecules in a competitive assay format. The ferrocene redox centers can be detected electrochemically. In order to facilitate the electron transfer, a gold layer structured in nanometer dimensions is used as an electrode. The functionalized silicon chip will be integrated into a micromachined fluid transport system. The resulting immunosensor device for estradiol represents a prototype allowing easy adaptation to other biotinylated analytes since the immobilized streptavidin serves as a universal anchor.

Bacterial Proteins↗