Molybdenum disulfide sphere-based electrochemical aptasensors for protein detection.

In this work, we report the development of an ultrasensitive sandwich-type electrochemical aptasensor for protein detection. The aptasensor is fabricated by using nitrogen-doped graphene oxide (N-GO) and Au nanoparticles (AuNPs) as sensing substrates, molybdenum disulfide (MoS2) spheres as the hybridization chain reaction (HCR) platform, and thrombin as the model protein. When the hybridization reaction is initiated through two biotinylated hairpin probes, vast horseradish peroxidases are immobilized on the long duplex by the biotin-avidin reaction. An electrochemical-chemical-chemical redox cycling reaction then takes place in the detection system, which contains p-dihydroxybenzene, ferrocene carboxylate and tris(2-carboxyethyl)phosphine. Benefiting from the good conductivity and high specific surface area of N-GO/AuNPs and MoS2 spheres, signal amplification of the HCR and detection system, and excellent selectivity of the aptamer and sandwich-type strategy, the proposed assay shows a wide linear range of 10 fM-0.1 nM towards thrombin with a detection limit of 27 aM (S/N = 3) along with clear distinction from different proteins. The proposed assay is successfully used to detect thrombin in human serum, which would have promising prospects for disease diagnosis and therapy.

Ultrasensitive electrochemical biosensing platform based on spherical silicon dioxide/molybdenum selenide nanohybrids and triggered Hybridization Chain Reaction.

An ultrasensitive sandwich-type electrochemical biosensor for DNA detection is developed based on spherical silicon dioxide/molybdenum selenide (SiO2@MoSe2) and graphene oxide-gold nanoparticles (GO-AuNPs) hybrids as carrier triggered Hybridization Chain Reaction (HCR) coupling with multi-signal amplification. The proposed sensoring assay utilizes a spherical SiO2@MoSe2/AuNPs as sensing platform and GO-AuNPs hybrids as carriers to supply vast binding sites. H2O2+HQ system is used for DNA detection and HCR as the signal and selectivity enhancer. The sensor is designed in sandwich type to increase the specificity. As a result, the present biosensor exhibits a good dynamic range from 0.1fM to 100pM with a low detection limit of 0.068fM (S/N=3). This work shows a considerable potential for quantitative detection of DNA in early clinical diagnostics.

Au nanoparticles/hollow molybdenum disulfide microcubes based biosensor for microRNA-21 detection coupled with duplex-specific nuclease and enzyme signal amplification.

An ultrasensitive electrochemical biosensor for detecting microRNAs is fabricated based on hollow molybdenum disulfide (MoS2) microcubes. Duplex-specific nuclease, enzyme and electrochemical-chemical-chemical redox cycling are used for signal amplification. Hollow MoS2 microcubes constructed by ultrathin nanosheets are synthesized by a facile template-assisted strategy and used as supporting substrate. For biosensor assembling, biotinylated ssDNA capture probes are first immobilized on Au nanoparticles (AuNPs)/MoS2 modified electrode in order to combine with streptavidin-conjugated alkaline phosphatase (SA-ALP). When capture probes hybridize with miRNAs, duplex-specific nuclease cleaves the formative duplexes. At the moment, the biotin group strips from the electrode surface and SA-ALP is incapacitated to attach onto electrode. Then, ascorbic acids induce the electrochemical-chemical-chemical redox cycling to produce electrochemical response in the presence of ferrocene methanol and tris (2-carboxyethyl) phosphine. Under optimum conditions, the proposed biosensor shows a good linear relationship between the current variation and logarithm of the microRNAs concentration ranging from 0.1fM to 0.1pM with a detection limit of 0.086fM (S/N=3). Furthermore, the biosensor is successfully applied to detect target miRNA-21 in human serum samples.

A layered tungsten disulfide/acetylene black composite based DNA biosensing platform coupled with hybridization chain reaction for signal amplification.

A 2-dimensional tungsten disulfide-acetylene black (WS2-AB) composite is synthesized by a simple hydrothermal method to achieve excellent electrochemical properties for applications as a DNA biosensor. The biosensor is fabricated based on the Au nanoparticles (AuNPs) and WS2-AB composite modified electrode, which subsequently is used to couple with a capture probe by an Au-S bond, then modified with target DNA, auxiliary DNA and bio-H1-bio-H2 (H1-H2) to perform hybridization chain reaction for signal amplification. Herein, two DNA hairpins H1 and H2 are opened by the recognition probe. The nicked double helices from hybridization chain reaction are used to immobilize horseradish peroxidase enzymes via biotin-avidin reaction, which produces signal-amplification detection of target DNA through the catalytic reaction of the hydrogenperoxide + hydroquinone system. Under optimum conditions, the as-prepared biosensor shows a good linear relationship between the current value and logarithm of the target DNA concentration ranging from 0.001 pM to 100 pM and a detection limit as low as 0.12 fM. Moreover, the fabricated biosensor exhibits good selectivity to differentiate the one-base mismatched DNA sequence. This work will open a pathway for ultrasensitive detection of other biorecognition events and gene-related diseases based on layered WS2-AB and hybridization chain reaction.

Ultrasensitive sensing platform for platelet-derived growth factor BB detection based on layered molybdenum selenide-graphene composites and Exonuclease III assisted signal amplification.

A highly sensitive and ultrasensitive electrochemical aptasensor for platelet-derived growth factor BB (PDGF-BB) detection is fabricated based on layered molybdenum selenide-graphene (MoSe2-Gr) composites and Exonuclease III (Exo III)-aided signal amplification. MoSe2-Gr is prepared by a simple hydrothermal method and used as a promising sensing platform. Exo III has a specifical exo-deoxyribonuclease activity for duplex DNAs in the direction from 3' to 5' terminus, however its activity is limited on the duplex DNAs with more than 4 mismatched terminal bases at 3' ends. Herein, aptamer and complementary DNA (cDNA) sequences are designed with four thymine bases on 3' ends. In the presence of target protein, the aptamer associates with it and facilitates the formation of duplex DNA between cDNA and signal DNA. The duplex DNA then is digested by Exo III and releases cDNA, which hybridizes with signal DNA to perform a new cleavage process. Nevertheless, in the absence of target protein, the aptamer hybridizes with cDNA will inhibit the Exo III-assisted nucleotides cleavage. The signal DNA then hybridizes with capture DNA on the electrode. Subsequently, horse radish peroxidase is fixed on electrode by avidin-biotin reaction and then catalyzes hydrogen peroxide and hydroquinone to produce electrochemical response. Therefore, a bridge can be established between the concentration of target protein and the degree of the attenuation of the obtained signal, providing a quantitative measure of target protein with a broad detection range of 0.0001-1 nM and a detection limit of 20 fM.

Ultrasensitive electrochemical sensing platform for microRNA based on tungsten oxide-graphene composites coupling with catalyzed hairpin assembly target recycling and enzyme signal amplification.

An ultrasensitive electrochemical biosensor for microRNA (miRNA) is developed based on tungsten oxide-graphene composites coupling with catalyzed hairpin assembly target recycling and enzyme signal amplification. WO3-Gr is prepared by a simple hydrothermal method and then coupled with gold nanoparticles to act as a sensing platform. The thiol-terminated capture probe H1 is immobilized on electrode through Au-S interaction. In the presence of target miRNA, H1 opens its hairpin structure by hybridization with target miRNA. This hybridization can be displaced from the structure by another stable biotinylated hairpin DNA (H2), and target miRNA is released back to the sample solution for next cycle. Thus, a large amount of H1-H2 duplex is produced after the cyclic process. At this point, a lot of signal indicators streptavidin-conjugated alkaline phosphatase (SA-ALP) are immobilized on the electrode by the specific binding of avidin-biotin. Then, thousands of ascorbic acid, which is the enzymatic product of ALP, induces the electrochemical-chemical-chemical redox cycling to produce a strongly electrochemical response in the presence of ferrocene methanol and tris (2-carboxyethyl) phosphine. Under the optimal experimental conditions, the established biosensor can detect target miRNA down to 0.05fM (S/N=3) with a linear range from 0.1fM to 100pM, and discriminate target miRNA from mismatched miRNA with a high selectivity.