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Biomedical subjects

Abdel-Nasser Kawde

Publications and source records attributed to Abdel-Nasser Kawde.

8 recordsLinked to original sources

Nanoparticle-based sensing of glycan-lectin interactions.

Here we present the first report on nanoparticle-based biosensing of glycan markers of diseases. The protocol relies on the competition between a nanocrystal (CdS)-tagged sugar and the target sugar for the binding sites of surface-confined lectin and monitoring the extent of competition through highly sensitive electrochemical detection of the captured nanocrystal. This development is expected to allow decentralized detection of carbohydrate moieties and lectin-carbohydrate interactions to be performed more rapidly, sensitively, inexpensively, and reliably.

Lectins↗

Sensitive and rapid electrochemical bioassay of glycosidase activity.

Here we present a highly sensitive, rapid and simple electrochemical assay for glycosidases based on treatment of the glycosidase with the appropriate p-nitrophenyl glycoside and anodic detection of released p-nitrophenol. The attractive characteristics of the new bioassay should facilitate advanced glycomic research and routine clinical diagnostics since glycosidases are associated with various diseases.

Bacteria↗

Quantum-dot/aptamer-based ultrasensitive multi-analyte electrochemical biosensor.

The coupling of aptamers with the coding and amplification features of inorganic nanocrystals is shown for the first time to offer a highly sensitive and selective simultaneous bioelectronic detection of several protein targets. This is accomplished in a single-step displacement assay in connection to a self-assembled monolayer of several thiolated aptamers conjugated to proteins carrying different inorganic nanocrystals. Electrochemical stripping detection of the nondisplaced nanocrystal tracers results in a remarkably low (attomole) detection limit, that is, significantly lower than those of existing aptamer biosensors. The new device offers great promise for measuring a large panel of disease markers present at ultralow levels during early stages of the disease progress.

Aptamers, Peptide↗

Carbon-nanotube-modified electrodes for amplified enzyme-based electrical detection of DNA hybridization.

Carbon-nanotube-modified glassy carbon (CNT/GC) transducers have been developed for enhancing the sensitivity and stability of enzyme-based electrochemical bioassays of DNA hybridization. The amplified signal reflects the interfacial accumulation of phenolic products of the alkaline-phosphatase tracer onto the CNT layer. In particular, chronopotentiometric measurements (following short accumulation periods) offer a substantial enhancement of the response of enzymatically liberated alpha-naphthol. The CNT modifier also leads to a dramatic improvement in the stability of the amperometric response of alpha-naphthol. These advantages of CNT/GC electrodes are illustrated from comparison to unmodified glassy carbon electrodes. Factors influencing the adsorptive accumulation of alpha-naphthol, and the overall performance of the new DNA assay, are assessed and optimized. The attractive performance characteristics of the new multi-amplification electrochemical detection of DNA hybridization are reported in connection to the detection of nucleic acid sequences related to the breast cancer BRCA1 gene.

Alkaline Phosphatase↗

Carbon-nanotube-modified glassy carbon electrodes for amplified label-free electrochemical detection of DNA hybridization.

The preparation and attractive performance of carbon-nanotube modified glassy-carbon (CNT/GC) electrodes for improved detection of purines, nucleic acids, and DNA hybridization are described. The surface-confined multiwall carbon-nanotube (MWCNT) facilitates the adsorptive accumulation of the guanine nucleobase and greatly enhances its oxidation signal. The advantages of CNT/GC electrodes are illustrated from comparison to the common unmodified glassy carbon, carbon paste and graphite pencil electrodes. The dramatic amplification of the guanine signal has been combined with a label-free electrical detection of DNA hybridization. Factors influencing the enhancement of the guanine signal are assessed and optimized. The performance characteristics of the amplified label-free electrochemical detection of DNA hybridization are reported in connection to measurements of nucleic-acid segments related to the breast-cancer BRCA1 gene.

Animals↗

Amplified label-free electrical detection of DNA hybridization.

A new protocol is described for amplifying label-free electrochemical measurements of DNA hybridization based on the enhanced accumulation of purine nucleobases in the presence of copper ions . Such electrical DNA assays involve hybridization of the target to inosine-substituted oligonucleotide probes (captured on magnetic beads), acidic dipurinization of the hybrid DNA, and adsorptive chronopotentiometric stripping measurements of the free nucleobases in the presence of copper ions. Both amplified adenine and guanine peaks can be used for detecting the DNA hybridization. The dramatic signal amplification advantage of this type of detection has been combined with efficient magnetic removal of non-complementary DNA, use of microliter sample volumes and disposable transducers. Factors influencing the signal enhancement were assessed and optimized. A detection limit of 40 fmol (250 pg) was obtained with 10 min hybridization and 5 min adsorptive-accumulation times. The advantages of this procedure were demonstrated by its application in the detection of DNA segments related to the BRCA1 breast cancer gene. The copper enhancement holds great promise not only for the detection of DNA hybridization, but also for trace measurement of nucleic acids.

Animals↗

Dual enzyme electrochemical coding for detecting DNA hybridization.

Enzyme-based hybridization assays for the simultaneous electrochemical measurements of two DNA targets are described. Two encoding enzymes, alkaline phosphatase and beta-galactosidase, are used to differentiate the signals of two DNA targets in connection to chronopotentiometric measurements of their electroactive phenol and alpha-naphthol products. These products yield well-defined and resolved peaks at +0.31 V (alpha-naphthol) and +0.63 V (phenol) at the graphite working electrode (vs. Ag/AgCl reference). The position and size of these peaks reflect the identity and level of the corresponding target. The dual target detection capability is coupled to the amplification feature of enzyme tags (to yield fmol detection limits) and with an efficient magnetic removal of non-hybridized nucleic acids. Proper attention is given to the choice of the substrates (for attaining well resolved peaks), to the activity of the enzymes (for obtaining similar sensitivities), and to the selection of the enzymes (for minimizing cross interferences). The new bioassay is illustrated for the simultaneous detection of two DNA sequences related to the BCRA1 breast-cancer gene in a single sample in connection to magnetic beads bearing the corresponding oligonucleotide probes. Prospects for electrochemical coding of multiple DNA targets are discussed.

Alkaline Phosphatase↗