Biomedical subjects

Damien Marchal

Publications and source records attributed to Damien Marchal.

6 recordsLinked to original sources

PubMed2007

Electrode surface confinement of self-assembled enzyme aggregates using magnetic nanoparticles and its application in bioelectrocatalysis.

Self-assembled enzyme aggregates, prepared from magnetic iron oxide nanoparticles, avidin, and a biotinylated redox enzyme, were shown particularly useful for the simple, fast, and efficient construction of highly enzyme-loaded electrodes with the help of a magnet. The approach was illustrated in the case of the bioelectrocatalytic oxidation of NADH by a diaphorase oxidoreductase in the presence of a ferrocene mediator. Two different self-assembling procedures were tested, taking advantage of the spontaneous aggregation of the nanoparticles in the presence of avidin and also of the multivalency binding of biotinylated diaphorase toward avidin. Activities of the bound and unbound diaphorase were systematically controlled allowing determination of the number of active biotinylated diaphorase per nanoparticle incorporated within each magnetic enzyme aggregate. An active enzyme loading capacity of up to 2.35 nmol mg-1 was found for the best nanostructured enzyme assembly, which is 200 times better than for commercialized magnetic micrometer-sized beads coated with streptavidin and saturated with diaphorase. With the help of a permanent magnet, the magnetic enzyme aggregates were finally magnetically collected as a film on the surface of a small screen-printed carbon electrode and the catalytic currents recorded by cyclic voltammetry. From the analysis of the steady-state catalytic current responses and the kinetic rate constants of biotinylated diaphorase, it was possible to determine the enzyme concentration within the magnetic films. Owing to the high enzyme loading in the aggregates of nanoparticles (i.e., 130 microM), the catalytic current responses were definitely higher than the ones measured at an electrode coated with a closed-packed monolayer of diaphorase or at an electrode covered with a film of magnetic micrometer-sized streptavidin beads saturated with diaphorase.

Avidin↗

PubMed2006

High amplification rates from the association of two enzymes confined within a nanometric layer immobilized on an electrode: modeling and illustrating example.

Electrochemical responses (e.g., chronoamperometric) obtained with an immobilized enzyme that produces an electroactive species may be used to quantitate the amount of enzyme or the concentration of its substrate. It is shown, on theoretical and experimental bases, that product-to-substrate coupling with a second enzyme co-immobilized with the first within one or within a small number of monolayers, allows high amplification rates (higher than 1000), avoids membrane transport limitations, and lends itself to precise kinetic analyses that provide guidelines for optimization of the analytical sensitivity. Very large amplification factors, as large as several thousands, can be reached experimentally, in agreement with appropriately derived theoretical predictions, thus opening the route to the rational design of high-performance substrate sensing or affinity assays applications.

Avidin↗

PubMed2006

Redox enzymes immobilized on electrodes with solution cosubstrates. General procedure for simulation of time-resolved catalytic responses.

In view of the existing and potential applications of electrochemical enzymatic catalysis with redox enzymes immobilized on the electrode surface in biosensors, a numerical calculation procedure for simulating their cyclic voltammetric responses is presented. It is applicable to systems involving a redox cosubstrate in solution. The cosubstrates, substrates, products, and inhibitors are assumed to diffuse linearly (planar electrode) between the electrode and the solution. The reactions in which the various forms of the immobilized enzyme participate may be as numerous and intricate as required by the simulation with no other restriction than the computing time. They may, at will, follow or not follow Michaelis-Menten kinetics. Slow charge-transfer cosubstrates are treated in the framework of Butler-Volmer kinetic law.

Catalysis↗

PubMed2006

Electrochemistry of immobilized redox enzymes: kinetic characteristics of NADH oxidation catalysis at diaphorase monolayers affinity immobilized on electrodes.

In the class of NADH:acceptor oxidoreductases, the diaphorase from Bacillus stearothermophilusis a particularly promising enzyme for sensing NADH, and indirectly a great number of analytes, when coupled with a NAD-dependent dehydrogenase as well as for the design of mono- and multienzyme affinity sensors. The design and rational optimization of such systems require devising immobilization procedures that prevent dramatic losses of the enzymatic activity and a full kinetic characterization of the immobilized enzyme system. Two immobilization procedures are described, which involve recognition of the biotinylated diaphorase by a monolayer of neutravidin adsorbed on the electrode surface either directly or through the intermediacy of a monolayer of biotinylated rabbit immunoglobulin. Thorough kinetic characterization of the two systems is derived from cyclic voltammetric responses. A precise estimate of the enzyme coverages is obtained after comparing the enzyme kinetics of the immobilized and the homogeneous system.

Biosensing Techniques↗

PubMed2004

Polymer-cushioned lipid bilayers in porous alumina.

Using small-angle neutron scattering (SANS) and cyclic voltammetry (CV), we show that model biological membranes can be deposited on a polymer cushion confined in highly regular porous alumina. The thicknesses of the dilute polymer cushion chemically bound to the alumina and of the supported bilayer are obtained for two polyethylene glycol cushions (PEG(5000) and PEG(20000)) and for a cushion made of chains bearing a lipid anchor at their free end (DSPE-PEG(3400)). The bilayers are studied well below and well above the chain melting temperature of the lipid mixture (DMPC/DMPE: 80/20), using a coenzyme (Ubiquinone, UQ(10)) as a redox probe for the voltammetry experiments. Analysis of the SANS form factor of the bilayers shows that the bilayer thickness can be extracted in this particular geometry. Using PEG chains grafted at a low surface density (D < 2R(g)), the thickness of the complete molecular construction is obtained by CV, which shows (after subtracting the bilayer thickness) that the polymer cushion thickness can be varied from 50 to 150 Angstroms. The values obtained with three different chain lengths, are in perfect agreement with the radius derived from the Flory theory.

Aluminum Oxide↗

PubMedDate not supplied

An intestinal surgery simulator: real-time collision processing and visualization.

This research work is aimed toward the development of a VR-based trainer for colon cancer removal. It enables the surgeons to interactively view and manipulate the concerned virtual organs as during a real surgery. First, we present a method for animating the small intestine and the mesentery (the tissue that connects it to the main vessels) in real-time, thus enabling user interaction through virtual surgical tools during the simulation. We present a stochastic approach for fast collision detection in highly deformable, self-colliding objects. A simple and efficient response to collisions is also introduced in order to reduce the overall animation complexity. Second, we describe a new method based on generalized cylinders for fast rendering of the intestine. An efficient curvature detection method, along with an adaptive sampling algorithm, is presented. This approach, while providing improved tessellation without the classical self-intersection problem, also allows for high-performance rendering thanks to the new 3D skinning feature available in recent GPUs. The rendering algorithm is also designed to ensure a guaranteed frame rate. Finally, we present the quantitative results of the simulations and describe the qualitative feedback obtained from the surgeons.

Algorithms↗