Recent Advances in Electrochemical Immunosensors.

Immunosensors have experienced a very significant growth in recent years, driven by the need for fast, sensitive, portable and easy-to-use devices to detect biomarkers for clinical diagnosis or to monitor organic pollutants in natural or industrial environments. Advances in the field of signal amplification using enzymatic reactions, nanomaterials such as carbon nanotubes, graphene and graphene derivatives, metallic nanoparticles (gold, silver, various oxides or metal complexes), or magnetic beads show how it is possible to improve collection, binding or transduction performances and reach the requirements for realistic clinical diagnostic or environmental control. This review presents these most recent advances; it focuses first on classical electrode substrates, then moves to carbon-based nanostructured ones including carbon nanotubes, graphene and other carbon materials, metal or metal-oxide nanoparticles, magnetic nanoparticles, dendrimers and, to finish, explore the use of ionic liquids. Analytical performances are systematically covered and compared, depending on the detection principle, but also from a chronological perspective, from 2012 to 2016 and early 2017.

Functionalization of single-walled carbon nanotubes for direct and selective electrochemical detection of DNA.

We report here a new strategy to graft both redox and DNA probes on carbon nanotubes to make a label-free DNA sensor. Oxidized single-walled carbon nanotubes are first immobilized on a self-assembled monolayer of cysteamine; then the redox probe, a quinone derivative 3-[(2-aminoethyl)sulfanyl-5-hydroxy-1,4-naphthoquinone], is grafted on the free carboxylic groups of the nanotubes. After that, for DNA probe grafting, new carboxylic sites are generated via an aryl diazonium route. After hybridization with a complementary sequence, the conformational changes of DNA could influence the redox kinetics of quinone, leading to a current increase of the redox signal, detected by square wave voltammetry. The system is selective, as it can discriminate a single mismatched sequence from the complementary one.

Hydroxynaphthoquinone ultrathin films obtained by diazonium electroreduction: toward design of biosensitive electroactive interfaces.

Electroactive 2-(phenylsulfanyl)-8-hydroxy-1,4-naphthoquinone has been electrodeposited via the reduction of the corresponding diazonium salt on Au electrodes. Surface characterizations by X-ray photoelectron spectroscopy (XPS) and infrared reflection-absorption spectroscopy (IRRAS) reveal that the mechanism of film deposition follows an aryl radical formation and its immobilization on the electrode surface. Electrochemical study shows that the surface coverage can be finely tuned (thickness between one and four layers) by adjusting the potential and the deposition time. By managing the potential applied when reducing diazonium in potentiostatic mode, the formed layer could mediate or not charge transfer. This is the first time that the films obtained by diazonium process are demonstrated to act as mediators in the growth process. Hence, with potentials higher than the formal potential of quinone group, very thin and homogeneous layers are obtained, whereas thicker films are formed when more cathodic potentials than that of quinone are applied. The possibility to manage the charge-transfer kinetics, the thickness, and the homogeneity of electroactive deposits is interesting in the scope of designing electrochemical transducers.

Nanometric layers for direct, signal-on, selective, and sensitive electrochemical detection of oligonucleotides hybridization.

We report a signal-on, reagentless electrochemical DNA biosensor, based on an electroactive self-assembled naphthoquinone derivative (JUG(thio)) monolayer. This system achieves highly sensitive (approximately 300 pM) and selective signal-on detection. Before hybridization, the single strand can interact with JUG(thio) and slow down the redox reaction. When the complementary target is added, the formation of the double helix eliminates the single strand/JUG(thio) interactions and the JUG(thio) redox rate, and hence the current increase.

Transistors for Chemical Monitoring of Living Cells.

We review here the chemical sensors for pH, glucose, lactate, and neurotransmitters, such as acetylcholine or glutamate, made of organic thin-film transistors (OTFTs), including organic electrochemical transistors (OECTs) and electrolyte-gated OFETs (EGOFETs), for the monitoring of cell activity. First, the various chemicals that are produced by living cells and are susceptible to be sensed in-situ in a cell culture medium are reviewed. Then, we discuss the various materials used to make the substrate onto which cells can be grown, as well as the materials used for making the transistors. The main part of this review discusses the up-to-date transistor architectures that have been described for cell monitoring to date.

Versatile transduction scheme based on electrolyte-gated organic field-effect transistor used as immunoassay readout system.

We report on an innovative heterogeneous bisphenol A (BPA) immunoassay based on an electrolyte-gated organic field-effect transistor whose organic semiconductor is poly(2,5-bis(3-tetradecylthiophen-2-yl)thieno[3,2-b]thiophene) co-crystallized with an alkyl derivative of bisphenol A. A decrease of the transistor output current is first observed upon antibody specific binding onto the organic semiconductor. Upon bisphenol A addition, the competitive dissociation of the antibody from the semiconductor surface leads to an opposite increase of the output current. We present here a proof-of-concept for bisphenol A detection; the device could be readily adapted to other small organic molecules of interest and is a promising tool for simple, low-cost, portable and easy-to-use biosensors.

Selectivity and sensitivity of a reagentless electrochemical DNA sensor studied by square wave voltammetry and fluorescence.

Poly(5-hydroxy-1,4-naphthoquinone-co-5-hydroxy-3-thioacetic acid-1,4-naphthoquinone)-modified electrode is used for the direct electrochemical detection of oligonucleotide hybridization. The polymer film presents well-defined electroactivity in the cathodic potential domain (between 0 and -0.8 V/SCE), due to the quinone group embedded into the polymer structure. The detection can be performed simply by square wave voltammetry. This sensor is a "signal-on" device and works with different oligonucleotide lengths, from 10 to 30 bases. Quantitative results from fluorescence are consistent with electrochemical data. It is confirmed that the signal increase in square wave voltammetry is unambiguously due to hybridization. The biosensor presents a detection limit of target of ca. 25 nM and is highly selective as it can discriminate single mismatch base.

Comparison of Electrochemical Immunosensors and Aptasensors for Detection of Small Organic Molecules in Environment, Food Safety, Clinical and Public Security.

We review here the most frequently reported targets among the electrochemical immunosensors and aptasensors: antibiotics, bisphenol A, cocaine, ochratoxin A and estradiol. In each case, the immobilization procedures are described as well as the transduction schemes and the limits of detection. It is shown that limits of detections are generally two to three orders of magnitude lower for immunosensors than for aptasensors, due to the highest affinities of antibodies. No significant progresses have been made to improve these affinities, but transduction schemes were improved instead, which lead to a regular improvement of the limit of detections corresponding to ca. five orders of magnitude over these last 10 years. These progresses depend on the target, however.

Quinone-based polymers for label-free and reagentless electrochemical immunosensors: application to proteins, antibodies and pesticides detection.

Polyquinone derivatives are widely recognized in the literature for their remarkable properties, their biocompatibility, simple synthesis, and easy bio-functionalization. We have shown that polyquinones present very stable electroactivity in neutral aqueous medium within the cathodic potential domain avoiding side oxidation of interfering species. Besides, they can act as immobilized redox transducers for probing biomolecular interactions in sensors. Our group has been working on devices based on such modified electrodes with a view to applications for proteins, antibodies and organic pollutants using a reagentless label-free electrochemical immunosensor format. Herein, these developments are briefly reviewed and put into perspective.

Copolythiophene-based water-gated organic field-effect transistors for biosensing.

This paper reports on the sensing of proteins using water-gated organic field-effect transistors. As a proof-of-concept, streptavidin and avidin were used, with a biotinylated polymer as the active sensing material. The latter is a copolythiophene modified to graft biotin by peptidic coupling. After characterization of its structure, it was integrated as the channel material into transistors and its interactions with several proteins were investigated. Non-specific interactions were reduced when the polymer surface was pretreated with 1-octanol. In this case, human serum albumin had no effect on the transistor characteristics whereas avidin and streptavidin led to a decrease of the drain current.

DNA electrochemical sensor based on conducting polymer: dependence of the "signal-on" detection on the probe sequence localization.

We show in this work that it is possible to make selective direct electrochemical hybridization detection of a target strand onto a probe strand immobilized on a conducting polymer modified with a quinone group, which presents cation-exchange properties. This leads to a "signal-on"detection, a unique behavior in comparison to similar systems described in the literature. It is shown that this system is efficient for various probe and target lengths (10-30 bp) and can discriminate a single mismatch. To go further in comprehension of the detection mechanism, a systematic study of the electrochemical response versus the probe sequence localization onto the immobilized strand is performed. For example, a 30-bp target strand is divided into three shorter 10-bp sequences (A-C, respectively), and we investigate the successive hybridization of these 1/3 strands onto the 30-bp probe strand. It is shown that one probe strand can be used to address several shorter targets.