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Biomedical subjects

Serge Cosnier

Publications and source records attributed to Serge Cosnier.

At least 19 recordsLinked to original sources
PubMed2006

Electrogenerated indium tin oxide-coated glass surface with photosensitive interfaces: surface analysis.

We present herein a photo-immobilization technique for the localized and specific conjugation of biochip platforms with different proteinaceous bioreceptors, such as antigen or antibodies. This methodology based on a photoactivable electrogenerated polymer film, pyrrole-benzophenone, allows the covalent immobilization of biomolecules through light mediation. The surface-conductive glass platform electropolymerized with poly(pyrrole-benzophenone) thin film may then be used to affinity-coat the chip with molecular recognition probes. This glass chip electroconductive surface modification is done by the deposition of a thin layer of indium tin oxide (ITO). Thereafter, pyrrole-benzophenone monomers are electropolymerized onto the conductive metal oxide surface and then exposed to an antigen Staphylococcal Enterotoxin B (SEB)) solution and illuminated with UV light (wavelength approximately 345 nm) through a mask. As a result of the photochemical reaction, a pattern thin layer of the antigen was covalently bound to the benzophenone-modified surface. Then the sample to be analyzed, along with its specific target antibody (anti-SEB antibodies), is introduced onto the glass surface and left to react with the previously photo-immobilized antigen. When the immuno-reaction is completed, the specifically attached immunoglobulin analytes are detected by using secondary antibodies conjugated with Fluorescein isothiocyanate (FITC). The fluorescence signal emanating from the biochip surface is then quantified by two methods, using a filtered intensified charge-coupled device (CCD) camera and a grating spectrometer.

Antibodies↗
PubMed2006

Laccase immobilization in redox active layered double hydroxides: a reagentless amperometric biosensor.

This paper describes a new system for amperometric determination of dissolved oxygen and its application for the detection of anionic toxic substances, which are known as enzyme inhibitors. This biosensor is based on the co-immobilization of laccase from Trametes versicolor and a redox active layered double hydroxide [Zn-Cr-ABTS] on a glassy carbon electrode. The electrochemical transduction step corresponds to the electrocatalytic reduction of O2 at 0.2V by laccase as catalyst and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) as mediator. Such device provides a fast and a sensitive response for dissolved oxygen determination between 6 x 10(-8) and 4 x 10(-6)M and very low detection limits for azide (5.5 nM), fluoride (6.9 nM) and cyanide (6.2 nM).

Biosensing Techniques↗
PubMed2006

Electroenzymatic polypyrrole-intercalator sensor for the determination of West Nile virus cDNA.

The chemical binding of a redox acridone derivative onto a polypyrrole film functionalized by N-hydroxysuccinimide groups provided an electrode capable of anchoring DNA duplex by simple insertion of the grafted acridone intercalator into the dsDNA solution. This electrode was applied for the detection of a ssDNA derived from a West Nile virus sequence. The latter was thus amperometrically detected after its hybridization in solution with a biotinylated complementary oligonucleotide followed by its anchoring and labeling by a glucose oxidase at 1 pg/mL.

Base Sequence↗
PubMed2006

Protease amperometric sensor.

An amperometric biosensor for the detection of trypsin was developed. The latter was based on a two-layer configuration, namely, a polymer-glucose oxidase inner layer and a gelatin outer layer. In the presence of glucose, the enzyme layer produces H2O2 and hence an amperometric signal due to H2O2 electrooxidation was generated by potentiostating the electrode at 0.6 V. The biosensor detects the change in the increase in the maximum current caused by the proteolytic digestion of gelatin, which covers the platinum electrodes, thereby facilitating a speedier access for the glucose substrate to the electrode modified with both poly(pyrrole-alkylammonium) and glucose oxidase molecules. Our biosensor detected low trypsin concentrations down to 42 pM with a response time of approximately 10 min, making it a very sensitive device in the detection of lower trypsin levels with such future putative applications as the diagnosis of pancreatic diseases.

Animals↗
PubMed2006

Development of amperometric biosensor for glucose based on a novel attractive enzyme immobilization matrix: calcium carbonate nanoparticles.

Calcium carbonate nanoparticles (nano-CaCO3) may be a promising material for enzyme immobilization owing to their high biocompatibility, large specific surface area and their aggregation properties. This attractive material was exploited for the mild immobilization of glucose oxidase (GOD) in order to develop glucose amperometric biosensor. The GOD/nano-CaCO3-based sensor exhibited a marked improvement in thermal stability compared to other glucose biosensors based on inorganic host matrixes. Amperometric detection of glucose was evaluated by holding the modified electrode at 0.60 V (versus SCE) in order to oxidize the hydrogen peroxide generated by the enzymatic reaction. The biosensor exhibited a rapid response (6s), a low detection limit (0.1 microM), a wide linear range of 0.001-12 mM, a high sensitivity (58.1 mAcm-2M-1), as well as a good operational and storage stability. In addition, optimization of the biosensor construction, the effects of the applied potential as well as common interfering compounds on the amperometric response of the sensor were investigated and discussed herein.

Biosensing Techniques↗
PubMed2006

Photoelectrochemical immunosensor for label-free detection and quantification of anti-cholera toxin antibody.

We demonstrate herein a newly developed photoelectrochemical immunosensor for the determination of anti-cholera toxin antibody by using a photosensitive biotinylated polypyrrole film. The latter was generated by electro-oxidation of a biotinylated tris(bipyridyl) ruthenium(II) complex bearing pyrrole groups. The photoexcitation of this modified electrode potentiostated at 0.5 V vs SCE, in the presence of an oxidative quencher, pentaaminechloro cobalt(III) chloride (15 mM), led to a cathodic photocurrent. As a result of the affinity interactions, a layer of biotinylated cholera toxin was firmly bound to the functionalized polypyrrole film via avidin bridges. The resulting modified electrodes were tested as immunosensors for the detection of the corresponding antibody from 0 to 200 microg mL(-)(1). The antibody concentration was measured through the decrease in photocurrent intensity resulting from its specific binding onto the polymeric coating, the detection limit being 0.5 microg mL(-)(1).

Antibodies↗
PubMed2006

Specific determination of As(V) by an acid phosphatase-polyphenol oxidase biosensor.

An original amperometric biosensor based on the simultaneous entrapment of acid phosphatase (AcP) and polyphenol oxidase (PPO) into anionic clays (layered double hydroxides) was developed for the specific detection of As(V). The functioning principle of the bienzyme electrode consisted of the successive hydrolysis of phenyl phosphate into phenol by AcP, followed by the oxidation of phenol into o-quinone by PPO. The phenyl phosphate concentration was, thus, monitored by potentiostating the biosensor at -0.2 V vs Ag/AgCl to detect amperometrically the generated quinone. The detection of As(V) was based on its inhibitory effect on AcP activity toward the hydrolysis of phenyl phosphate into phenol. The As(V) can be specifically determined in pH 6.0 acetate buffer without any interferences of As(III) or phosphate, the detection limit being 2 nM or 0.15 ppb after an incubation step for 20 min.

Acid Phosphatase↗
PubMed2006

Amperometric phenol biosensor based on laponite clay-chitosan nanocomposite matrix.

A novel strategy to fabricate an amperometric biosensor for phenol determination based on chitosan/laponite nanocomposite matrix was described. The composite film was used to immobilize PPO on the surface of a glassy carbon electrode. Chitosan was utilized to improve the analytical performance of the pure clay-modified bioelectrode. The biosensor exhibited a series of properties: good affinity to its substrate (the apparent Michaelis-Menten constant for the sensor was found to be 0.16 mM), high sensitivity (674 mA M(-1)cm(-2) for catechol) and remarkable long-term stability in storage (it retains 88% of the original activity after 60 days). In addition, optimization of the biosensor construction as well as effects of experimental variables such as pH, operating potential and temperature on the amperometric response of the sensor were discussed.

Adsorption↗
PubMed2005

Electrogeneration of a poly(pyrrole)-NTA chelator film for a reversible oriented immobilization of histidine-tagged proteins.

This contribution reports, for the first time, the synthesis and electropolymerization of a pyrrole N-substituted by a nitrilotriacetic acid acting as a chelating center of Cu2+. A step-by-step approach for protein immobilization was developed via the successive coordination of Cu2+ and histidine-tagged proteins. The self-assembly of histidine-tagged glucose oxidase led to the formation of a close-packed enzyme monolayer at the poly(pyrrole) surface, and the reversibility and reproducibility of this affinity process were demonstrated.

Avidin↗
PubMed2004

Construction of amperometric immunosensors based on the electrogeneration of a permeable biotinylated polypyrrole film.

The construction of amperometric immunosensors to cholera antitoxin immunoglobulins were shown to have improved sensitivity when the cholera toxin B subunit biorecognition entity was linked to an electrogenerated biotinylated polypyrrole film copolymerized with pyrrole-lactobionamide monomer. The copolymer exhibits greater film permeability than biotinylated polypyrrolic or polyphenolic films for the permeation of electroactive species. Hence, when the presence of the HRP marker of the immunoassay was determined using hydroquinone, the production of electroactive quinone was shown to permeate faster to the electrode, thus providing a faster response time.

Antibodies↗
PubMed2004

HRP/[Zn-Cr-ABTS] redox clay-based biosensor: design and optimization for cyanide detection.

A novel inexpensive and simple amperometric biosensor, based on the immobilization of HRP into redox active [Zn-Cr-ABTS] layered double hydroxide, is applied to the determination of cyanide. The electrochemical transduction step corresponds to the reduction at 0.0 V of ABTS+* enzymatically formed in the presence of H2O2. The biosensor has a fast response to H2O2 (8s) with a linear range of 1.7 x 10(-9) to 2.1 x 10(-6) M and a sensitivity of 875 mA M(-1) cm(-2). The apparent Michaelis-Menten constant (KMapp) is 12 microM. The detection of cyanide is performed via its non competitive inhibiting action on the HRP/[Zn-Cr-ABTS] electrode. The concentration range of the linear response and the apparent inhibition constant (ki) are 5 x 10(-9) to 4 x 10(-8) and 1.4 x 10 (-7) M, respectively.

Aluminum Silicates↗
PubMed2004

Subnanomolar cyanide detection at polyphenol oxidase/clay biosensors.

A novel, inexpensive, and simple amperometric biosensor based on immobilization of polyphenol oxidase (PPO) into Zn-Al layered double hydroxides, also called anionic clays, is applied for determination of cyanide. The detection of cyanide was performed via its inhibiting action on the PPO electrode. Measurement was carried out with 3,4-dihydroxyphenylacetic acid as enzyme substrate, the enzymatically generated quinoid products being electroreduced at -0.2 V. An extremely sensitive detection limit (0.1 nM) was obtained for cyanide. Enzyme immobilization into an anionic exchanger clay seems to cause an increase in cyanide inhibition effects because of anion accumulation in the clay matrix.

Biosensing Techniques↗
PubMed2004

Amperometric detection of phenolic compounds by polypyrrole-based composite carbon paste electrodes.

This contribution describes new composite carbon paste electrodes (CPEs) for the determination of phenolic compounds. The composite CPEs were prepared by in situ generation of polypyrrole (PPy) within a paste containing the enzyme polyphenol oxidase (PPO). The best paste composition (enzyme/pyrrole monomer/carbon particles/Nujol) was determined for a model enzyme, glucose oxidase (GOx) according to the enzymatic activity of the resulting electrodes and to the enzyme leakage from the paste during storage in phosphate buffer. The in situ electrogenerated PPy improves the enzyme immobilisation within the paste since practically no enzyme was lost in solution after 72 h of immersion. Moreover, the enzyme activity remains particularly stable under storage since the biocomposite structure conserves 80% of its activity after 1 month of storage. Following the optimisation of the paste composition, PPO-based carbon paste biosensors were prepared and presented excellent analytical properties toward catechol detection with a sensitivity of 4.7 A M(-1) cm(-2) and a response time lower than 20 s. The resulting biosensors were applied to the determination of polyphenolic compounds (e.g., epicatechin and ferulic acid).

Biosensing Techniques↗
PubMed2003

Biosensors based on electropolymerized films: new trends.

Electropolymerized films have received considerable attention in the development of biosensors and biochips, and are advancing rapidly. This paper reviews recent advances and scientific progress in electrochemical immobilization procedures for biological macromolecules on electrodes via electrogenerated polymer films. Biomolecule immobilization is classified as covalent linkage, attachment by affinity interactions, and physical entrapment. The last approach entails the use of conducting and non-conducting films, composite polymer films, and templates for the electropolymerization process. Some advances in the electrochemical transduction of biological events (enzymatic reaction, immunoreaction, or oligonucleotide hybridization) involving the redox properties or the conductivity of electropolymerized films are also presented.

Biosensing Techniques↗