Mohamed Braiek

An Electrochemical Immunosensor for Detection of Staphylococcus aureus Bacteria Based on Immobilization of Antibodies on Self-Assembled Monolayers-Functionalized Gold Electrode

The detection of pathogenic bacteria remains a challenge for the struggle against biological weapons, nosocomial diseases, and for food safety. In this research, our aim was to develop an easy-to-use electrochemical immunosensor for the detection of pathogenic Staphylococcus aureus ATCC25923. The biosensor was elaborated by the immobilization of anti-S. aureus antibodies using a self-assembled monolayer (SAMs) of 3-Mercaptopropionic acid (MPA). These molecular assemblies were spontaneously formed by the immersion of the substrate in an organic solvent containing the SAMs that can covalently bond to the gold surface. The functionalization of the immunosensor was characterized using two electrochemical techniques: cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Here, the analysis was performed in phosphate buffer with ferro/ferricyanide as the redox probe. The EIS technique was used for affinity assays: antibody-cell binding. A linear relationship between the increment in the electron transfer resistance (RCT) and the logarithmic value of S. aureus concentration was observed between 10 and 106 CFU/mL. The limit of detection (LOD) was observed at 10 CFU/mL, and the reproducibility was calculated to 8%. Finally, a good selectivity versus E. coli and S. epidermidis was obtained for our developed immunosensor demonstrating its specificity towards only S. aureus.

Aflatoxin B1 Detection Using a Highly-Sensitive Molecularly-Imprinted Electrochemical Sensor Based on an Electropolymerized Metal Organic Framework

A sensitive electrochemical molecularly-imprinted sensor was developed for the detection of aflatoxin B1 (AFB1), by electropolymerization of p-aminothiophenol-functionalized gold nanoparticles in the presence of AFB1 as a template molecule. The extraction of the template leads to the formation of cavities that are able to specifically recognize and bind AFB1 through π-π interactions between AFB1 molecules and aniline moities. The performance of the developed sensor for the detection of AFB1 was investigated by linear sweep voltammetry using a hexacyanoferrate/hexacyanoferrite solution as a redox probe, the electron transfer rate increasing when the concentration of AFB1 increases, due to a p-doping effect. The molecularly-imprinted sensor exhibits a broad linear range, between 3.2 fM and 3.2 µM, and a quantification limit of 3 fM. Compared to the non-imprinted sensor, the imprinting factor was found to be 10. Selectivity studies were also performed towards the binding of other aflatoxins and ochratoxin A, proving good selectivity.

Recent Advances in Electrospun Nanofiber Interfaces for Biosensing Devices

Electrospinning has emerged as a very powerful method combining efficiency, versatility and low cost to elaborate scalable ordered and complex nanofibrous assemblies from a rich variety of polymers. Electrospun nanofibers have demonstrated high potential for a wide spectrum of applications, including drug delivery, tissue engineering, energy conversion and storage, or physical and chemical sensors. The number of works related to biosensing devices integrating electrospun nanofibers has also increased substantially over the last decade. This review provides an overview of the current research activities and new trends in the field. Retaining the bioreceptor functionality is one of the main challenges associated with the production of nanofiber-based biosensing interfaces. The bioreceptors can be immobilized using various strategies, depending on the physical and chemical characteristics of both bioreceptors and nanofiber scaffolds, and on their interfacial interactions. The production of nanobiocomposites constituted by carbon, metal oxide or polymer electrospun nanofibers integrating bioreceptors and conductive nanomaterials (e.g., carbon nanotubes, metal nanoparticles) has been one of the major trends in the last few years. The use of electrospun nanofibers in ELISA-type bioassays, lab-on-a-chip and paper-based point-of-care devices is also highly promising. After a short and general description of electrospinning process, the different strategies to produce electrospun nanofiber biosensing interfaces are discussed.

An Immunosensor for Pathogenic Staphylococcus aureus Based on Antibody Modified Aminophenyl-Au Electrode

The objective of this work is to elaborate an immunosensing system which will detect and quantify Staphylococcus aureus bacteria. A gold electrode was modified by electrografting of 4-nitrophenyl diazonium, in situ synthesized in acidic aqueous solution. The immunosensor was fabricated by immobilizing affinity-purified polyclonal anti S. aureus antibodies on the modified gold electrode. Cyclic voltammetry (CV) and Faradaic Electrochemical Impedance Spectroscopy (EIS) were employed to characterize the stepwise assembly of the immunosensor. The performance of the developed immunosensor was evaluated by monitoring the electron-transfer resistance detected using Faradaic EIS. The experimental results indicated a linear relationship between the relative variation of the electron transfer resistance and the logarithmic value of S. aureus concentration, with a slope of 0.40 ± 0.08 per decade of concentration. A low quantification limit of CFU per ml and a linear range up to CFU per mL were obtained. The developed immunosensors showed high selectivity to Escherichia coli and Staphylococcus saprophyticus.

An enzyme biosensor based on beta-galactosidase inhibition for electrochemical detection of cadmium (II) and chromium (VI)

ABSTRACT The focus of this article is the development and optimisation of a simple, sensitive, selective biosensor for the detection of heavy metals, through inhibition of β-galactosidase (β-gal) from Aspergillus oryzae after its immobilisation on an electrochemical transducer by cross-linking with glutaraldehyde. The inhibition of β-gal by Cd(II) and Cr(VI) was demonstrated by the decrease of β-gal activity monitored by the conductometric signal. Under optimal conditions, the developed voltammetric and impedimetric biosensor, based on the specific interaction of metallic ions with beta-galactosidase that increases the electron transfer of the redox probe, presented a wide range of linearity. Detection limits of 3.12 × 10–10 M (91.7 ng/L) were obtained, using both techniques, for Cr(VI). For Cd(II) detection limits were 2.85 × 10–8 M (6.95 µg/L) using EIS and 3.22 × 10–11 M (7.61 ng/L) using square wave voltammetry (SWV).

A new sensitive and selective sensor for heavy metal ions based on tannin extracted from the skin of Punica granatum L

ABSTRACT This article presents the development of a sensor made from a gold electrode and a receiving polymeric membrane based on tannin extracted from the skin of Punica granatum. L (pomegranate) for real-time detection of heavy metals in a hydrous environment. The basic principle of this device is the complexing (chelating) of metal ions through the adjacent hydroxyl groups contained in the chemical structures of the tannins. The electrochemical characterisation was performed by using electrochemical impedance spectroscopy and square wave voltammetry. Other morphological and structural analyses were performed by using Fourier transform infrared spectroscopy and atomic-force microscopy. The results obtained showed the high sensitivity of the developed device (detection limits of 6.35 × 10−9 g L−1 for Cu2+, 1.1 × 10−8 g L−1 for Cd2+ and 2 × 10−7 g L−1 for Pb2+) and the possibility of simultaneously detecting several heavy metals, each one in a highly selective manner with highly acceptable response time (48s).

Effect of Perfluorinated-Hexaethylene Glycol Functionalization of Gold Nanoparticles on the Enhancement of the Response of an Enzymatic Conductometric Biosensor for Urea Detection

In conductometric enzymatic biosensors, enzymatic reaction is confined close to the interdigitated electrode surface, because enzyme is cross-linked in contact with this surface in the presence or absence of nanoparticles. The effect of the use of a new type of doubly-functionalized gold nanoparticles (PF-HEG-Au NPs) on the response of conductometric biosensor based on interdigitated electrodes (IDEs), for the detection of enzymatic substrates was studied. Gold nanoparticles (AuNPs) were first synthesized following the citrate process, with an average diameter of 14 nm. AuNPs were then functionalized with 11-mercaptoundecylhexaethyleneglycol (HEG) and then with 1H,1H,2H,2H-perfluorodecanethiol (PF). The doubly-functionalized AuNPs were characterized using TEM, UV-Vis spectrophotometry and FTIR spectroscopy. Urease, mixed with these doubly functionalized AuNPs, was then cross-linked with glutaraldhedyde vapor on the IDE surface. In the presence of urea, the conductometric response was measured in a differential mode. The best sensitivities for urea detection were obtained with PF-HEG-Au NPs (520 µS /mM and 0.5µM of detection limit), as compared to 284µS/mM and 2µM of detection limit with bare Au NPs, PF-AuNPs and HEG-AuNPs, and 1.07µS/mM and 100 µM of detection limit with urease directly crosslinked on IDEs.When stored in phosphate buffer (5 mM, pH 6.7) at 4 °C, the biosensor with PF-HEG-Au NPs showed good stability for more than 12 days.

5.1.1 Detection of pathogenic Staphylococcus aureus bacteria by Electrochemical Impedance

The timely detection of pathogens is a subject of great importance. In this work, the objective is to elaborate an immunosensing system for detection and quantification of Staphylococcus aureus. Poyclonal anti-S. aureus are immobilized onto gold electrodes via chemical bond formation between antibody amino groups and a carboxylic acid containing self-assembled molecular monolayer. The evaluation of the developed immunosensor performance was accomplished through the monitoring of the electron-transfer resistance detected by electrochemical impedance spectroscopy in the presence of [Fe(CN)6 3]/[Fe(CN)6 4] as redox probe. A low detection limit of 10 fcu/ml and a linear range up to 10 7