Electrochemiluminescence Aptasensor Research Articles

Efficient electrochemiluminescence quenching of carbon-coated petalous CdS nanoparticles for an ultrasensitive tumor marker assay through coreactant consumption by G-quadruplex-hemin decorated Au nanorods

A novel competitive electrochemiluminescence aptasensor was designed for the detection of carcinoembryonic antigen using carbon-coated petalous CdS nanopaticles as an ECL emitter and DNAzyme/Au nanorods–complementary DNA as a quenching probe.

Ratiometric electrochemiluminescent aptasensor array for antibiotic based on internal standard method and spatial-resolved technique

A novel ratiometric electrochemiluminescent (R-ECL) aptasensor array was fabricated on a homemade screen-printed carbon electrode (SPCE) for antibiotic chloramphenicol (CAP) based on the ratio of working signal to internal reference signal. The SPCE substrate consists of a common Ag/AgCl reference electrode, carbon counter electrode and two spatial-resolved carbon working electrodes (WE1 and WE2). In the system, luminol-gold nanoparticles (L-Au NPs) as working signal tags were modified on WE1, and CdS quantum dots (QDs) were modified on WE2 as internal reference to provide a correction for avoiding the environmental effects. The L-Au NPs at WE1 were functionalized with a CAP aptamer′s DNA complementary (DNA1) and then CAP aptamer (DNA2) that was tagged with chlorogenic acid (CA) as quenchers of L-Au NPs. Upon CAP adding to WE1, the DNA2-CAP complexes were formed due to the highly affinity of CAP towards DNA2. Meanwhile, it could release the CA, which recovered the ECL of L-Au NPs. This ratio of ECL signals for quantification between target probes (at WE1) to internal standard probes (at WE2) could reduce the matrix environmental interference to a certain degree. Moreover, the two probes immobilized on two working electrodes while not one electrode, could also reduce cross-talk interference between each other. Such R-ECL approach offers a selective and sensitive platform and shows a great utility for detection of CAP in the food analysis.

A regenerable electrochemiluminescence aptasensor incorporating poly(ethylenimine) and thiosemicarbazide as dual co-reactants for signal amplification

A regenerable electrochemiluminescence aptasensor for thrombin (TB) has been developed using dual co-reactants of poly(ethylenimine) (PEI) and thiosemicarbazide (TSC) for signal amplification. Herein, Au nanoparticle (AuNPs) were in-situ electrodeposited on glassy carbon electrode (GCE) in advance, and then the colloidal gold nanoparticle (nano-Au) and TSC were alternately self-assembled on AuNPs/GCE to obtain nano-Au/TSC/AuNPs multi-layers for 5’-thiolated partly complementary strand (PCS) immobilization. Subsequently, the multifunctional binding aptamer (MBA), consisted of an adenosine triphosphate (ATP) binding aptamer (ABA) and a thrombin binding aptamer (TBA), was hybridized with PCS to generate double-strands DNA (PCS/MBA). Meanwhile, the signal probe was fabricated by immobilizing detection thrombin aptamer (TBA 2) on the surface of PEI enhanced Ru(bpy)32+ doped nanospheres. On the basis of sandwich-type tactics, TB was measured quantitatively in the range from 1×10−5nM to 0.1nM. More importantly, the aptasensor could regenerate by treating with ATP to take off MBA based on ATP-induced conformation change, leaving only PCS onto the modified electrode to re-hybridize with MBA. The proposed aptasensor exhibited good selectivity, acceptable stability and satisfactory regenerability.

New Signal Amplification Strategy Using Semicarbazide as Co-reaction Accelerator for Highly Sensitive Electrochemiluminescent Aptasensor Construction

A highly sensitive electrochemiluminescent (ECL) aptasensor was constructed using semicarbazide (Sem) as co-reaction accelerator to promote the ECL reaction rate of CdTe quantum dots (CdTe QDs) and the co-reactant of peroxydisulfate (S2O8(2-)) for boosting signal amplification. The co-reaction accelerator is a species that when it is introduced into the ECL system containing luminophore and co-reactant, it can interact with co-reactant rather than luminophore to promote the ECL reaction rate of luminophore and co-reactant; thus the ECL signal is significantly amplified in comparison with that in which only luminophore and co-reactant are present. In this work, the ECL signal probes were first fabricated by alternately assembling the Sem and Au nanoparticles (AuNPs) onto the surfaces of hollow Au nanocages (AuNCs) via Au-N bond to obtain the multilayered nanomaterials of (AuNPs-Sem)n-AuNCs for immobilizing amino-terminated detection aptamer of thrombin (TBA2). Notably, the Sem with two -NH2 terminal groups could not only serve as cross-linking reagent to assemble AuNPs and AuNCs but also act as co-reaction accelerator to enhance the ECL reaction rate of CdTe QDs and S2O8(2-) for signal amplification. With the sandwich-type format, TBA2 signal probes could be trapped on the CdTe QD-based sensing interface in the presence of thrombin (TB) to achieve a considerably enhanced ECL signal in S2O8(2-) solution. As a result, the Sem in the TBA2 signal probes could accelerate the reduction of S2O8(2-) to produce the more oxidant mediators of SO4(•-), which further boosted the production of excited states of CdTe QDs to emit light. With the employment of the novel co-reaction accelerator Sem, the proposed ECL biosensor exhibited ultrahigh sensitivity to quantify the concentration of TB from 1 × 10(-7) to 1 nM with a detection limit of 0.03 fM, which demonstrated that the co-reaction accelerator could provide a simple, efficient, and low-cost approach for signal amplification and hold great potential for other ECL biosensors construction.

Label-Free Electrochemiluminescence Aptasensor for 2,4,6-Trinitrotoluene Based on Bilayer Structure of Luminescence Functionalized Graphene Hybrids.

The electrochemiluminescence (ECL) behavior of N-(aminobutyl)-N-(ethylisoluminol)/hemin dual-functionalized graphene hybrids (A-H-GNs) and luminol-functionalized silver/graphene oxide composite (luminol-AgNPs-GO) was investigated under cyclic voltammetry and pulse potential. It was found that A-H-GNs and luminol-AgNPs-GO exhibited excellent ECL activity. On this basis, a label-free ECL aptasensor for 2,4,6-trinitrotoluene (TNT) detection was developed based on bilayer structure of luminescence functionalized graphene hybrids consisting of A-H-GNs and luminol-AgNPs-GO. First, positively charged chitosan-coated A-H-GNs were modified on the surface of indium-doped tin oxide electrode by simple dripping and drying in the air; after that, the modified electrode was immersed in negatively charged luminol-AgNPs-GO modified with aptamer (apta-biotin-SA-luminol-AgNPs-GO) to form apta-biotin-SA-luminol-AgNPs-GO/CS-A-H-GNs/ITO electrode (i.e., aptasensor) by electrostatic interaction. In the presence of TNT, a remarkable decrease in ECL signals was observed due to the formation of aptamer-TNT complex. TNT could be detected based on the inhibition effect. The aptasensor exhibits a wide dynamic range from 1.0 × 10(-12) to 1.0 × 10(-9) g/mL, with a low detection limit of 6.3 × 10(-13) g/mL for the determination of TNT, which is superior to most previously reported bioassays for TNT. Moreover, the proposed aptasensor has been successfully applied to the detection of TNT in environmental water. It is sensitive, selective, and simple, avoiding complicated labeling and purification procedures. Due to the wide target recognition range of aptamer, this strategy provides a promising way to develop new aptasensor for other analytes.

The Ru complex and hollow gold nanoparticles branched-hydrogel as signal probe for construction of electrochemiluminescent aptasensor.

In this work, a novel Ru complex and hollow gold nanoparticles branched-poly(N-(3-aminopropyl)methacrylamide) hydrogel composites (pNAMA-Ru-HGNPs) were prepared and used as electrogenerated chemiluminescence (ECL) signal probe to construct aptasensor for ultrasensitive detection of thrombin (TB). Herein, [Ru(phen)2(cpaphen)](2+) linked N-(3-aminopropyl)methacrylamide and hollow gold nanoparticles functionalized N-(3-aminopropyl)methacrylamide were used as two polymer monomers to prepare pNAMA-Ru-HGNPs composites via free-radical polymerization. The obtained hydrogel composite, containing amount of Ru complex and HGNPs, were used as effective tag-carriers for the immobilization of thrombin binding aptamer II (TBA II) to form the pNAMA-Ru-HGNPs labeled TBA II (pNAMA-Ru-HGNPs-TBA II). For building the interface of the aptasensor, dendritic gold nanoparticles reduced by poly(ethyleneimine) (PEI@DGNPs) were modified on the carbon nanotube-nafion (CNTs-Nf) coated electrode through electrostatic adsorption, which was used not only as matrix for immobilization of thrombin binding aptamer I (TBA I) but also as enhancer to amplify the ECL signal because PEI is an efficient co-reactant of Ru complex. Target TB was sandwiched between pNAMA-Ru-HGNPs-TBA II and TBA I, resulting in the ECL signals relevant to the TB concentrations. Combining the novel pNAMA-Ru-HGNPs containing amount of Ru complex as the ECL signal probe and PEI@DGNPs as the enhancer for signal amplification, the sandwich ECL aptasensor was constructed for the detection of TB with a wide range of 1.0 fM to 10 pM and a low detection of 0.54 fM.

Triple Quenching of a Novel Self-Enhanced Ru(II) Complex by Hemin/G-Quadruplex DNAzymes and Its Potential Application to Quantitative Protein Detection.

Herein, a novel "on-off" electrochemiluminescence (ECL) aptasensor for highly sensitive determination of thrombin has been constructed based on the triple quenching of the effect of hemin/G-quadruplex DNAzymes upon the Ru(II) complex-based ECL system. First, a strong initial ECL signal was achieved by the dual amplification strategies of (i) intramolecular coreaction of a self-enhanced Ru(II)-based molecule (PTCA-PEI-Ru(II)) and (ii) intermolecular coreaction between PTCA-PEI-Ru(II) and nicotinamide adenine dinucleotide (NADH), which was named the signal-on state. Then, a novel triple quenching of the effect of multifunctional hemin/G-quadruplex DNAzymes upon the Ru(II) complex-based ECL system was designed to realize the desirable signal-off state, which was outlined as follows: (i) the hemin/G-quadruplex DNAzymes mimicked NADH oxidase to oxidize NADH and in situ generate the H2O2, consuming the coreactant of NADH; (ii) its active center of hemin could oxidize the excited state PTCA-PEI-Ru(II)* to PTCA-PEI-Ru(III), making the energy and electron transfer quench; (iii) it also acted as horseradish peroxidase (HRP) to catalyze the H2O2 for in situ producing the quencher of O2. Based on triple quenching of the effect of hemin/G-quadruplex DNAzymes, the highly sensitive "on-off" thrombin aptasensor was developed with a wide linear detection range of 1.0 × 10(-14) M to 1.0 × 10(-10) M and a detection limit down to the femtomolar level.

Petal-like CdS nanospheres-based electrochemiluminescence aptasensor for detection of IgE with gold nanoparticles amplification

A facile label-free electrochemiluminescence (ECL) aptasensor was designed for sensitive detection of human immunoglobulin E (IgE) using petal-like CdS nanospheres and Au nanoparticles (AuNPs) as sensing platform. To construct the aptasensor, petal-like CdS nanospheres as ECL emitter were firstly synthesized and immobilized on the chitosan-coated glassy carbon electrode (GCE) surface. Chitosan was coated on the CdS/CS/GCE again with two coating numbers to produce a stable ECL signal and facilitate subsequent AuNPs immobilization. The construction of aptasensor was achieved after IgE aptamer was adsorbed onto the AuNPs. The detection of IgE was performed upon the incubation of the interface with target protein IgE. Under the optimum conditions, the ECL signal decreased depending linearly on the logarithmic value of IgE concentration ranging from 5.0×10−13 to 1.0×10−9M with a regression equation of I=−15254.8−2129.3 logc (R2=0.996). The detection limit was experimentally found to be 8.0×10−14M. The applicability of the constructed aptasensor was demonstrated in the determination of IgE in human serum samples.

A novel "dual-potential" electrochemiluminescence aptasensor array using CdS quantum dots and luminol-gold nanoparticles as labels for simultaneous detection of malachite green and chloramphenicol.

A novel type of "dual-potential" electrochemiluminescence (ECL) aptasensor array was fabricated on a homemade screen-printed carbon electrode (SPCE) for simultaneous detection of malachite green (MG) and chloramphenicol (CAP) in one single assay. The SPCE substrate consisted of a common Ag/AgCl reference electrode, carbon counter electrode and two carbon working electrodes (WE1 and WE2). In the system, CdS quantum dots (QDs) were modified on WE1 as cathode ECL emitters and luminol-gold nanoparticles (L-Au NPs) were modified on WE2 as anode ECL emitters. Then the MG aptamer complementary strand (MG cDNA) and CAP aptamer complementary strand (CAP cDNA) were attached on CdS QDs and L-Au NPs, respectively. The cDNA would hybridize with corresponding aptamer that was respectively tagged with cyanine dye (Cy5) (as quenchers of CdS QDs) and chlorogenic acid (CA) (as quenchers of l-Au NPs) using poly(ethylenimine) (PEI) as a bridging agent. PEI could lead to a large number of quenchers on the aptamer, which increased the quenching efficiency. Upon MG and CAP adding, the targets could induce strand release due to the highly affinity of analytes toward aptamers. Meanwhile, it could release the Cy5 and CA, which recovered cathode ECL of CdS QDs and anode ECL of L-Au NPs simultaneously. This "dual-potential" ECL strategy could be used to detect MG and CAP with the linear ranges of 0.1-100 nM and 0.2-150 nM, with detection limits of 0.03 nM and 0.07 nM (at 3sB), respectively. More importantly, this designed method was successfully applied to determine MG and CAP in real fish samples and held great potential in the food analysis.

An electrochemiluminescence aptasensor based on flowerlike CdS–MoS2 composites and DNAzyme for detection of immunoglobulin E

A novel electrochemiluminescence (ECL) aptasensor based on CdS–MoS2 nanocomposites was designed for detection of immunoglobulin E (IgE). The flowerlike CdS–MoS2 composites were synthesized through a simple one-pot hydrothermal method. The CdS–MoS2 composites were immobilized on glassy carbon electrode surface as ECL emitter attached a complementary ssDNA (NH2–ssDNA) of IgE. IgE aptamer (T-Apt) with DNAzyme–AuNPs can hybridize with NH2–ssDNA to form dsDNA. The DNAzyme with horseradish peroxidase-like activity could electrocatalyze the reduction of H2O2, the coreactant of CdS–MoS2 ECL emission, leading to an obvious ECL quenching. In the presence of IgE, IgE conjugated with DNAzyme–AuNPs–T-Apt and thereafter the conjugations released, which decreased the consumption of H2O2 by DNAzyme–AuNPs, produced an increased ECL signal. The changed ECL intensity is proportional to concentration of IgE in the range of 0.001–10.0nM with the detection limit of 0.34pM (S/N=3). The prepared aptasensor has been applied to determination of IgE in human serum samples. The recoveries of IgE in human serum samples are between 83.0% and 108% with the RSD values no more than 3.8%. The developed sensor with CdS–MoS2 composites and DNAzyme features high sensitivity, good selectivity and wide linear range, suggesting the potential applications in biochemical analysis.

Application of Europium Multiwalled Carbon Nanotubes as Novel Luminophores in an Electrochemiluminescent Aptasensor for Thrombin Using Multiple Amplification Strategies.

A novel electrochemiluminescent (ECL) aptasensor was proposed for the determination of thrombin (TB) using exonuclease-catalyzed target recycling and hybridization chain reaction (HCR) to amplify the signal. The capture probe was immobilized on an Au-GS-modified electrode through a Au-S bond. Subsequently, the hybrid between the capture probe and the complementary thrombin binding aptamer (TBA) was aimed at obtaining double-stranded DNA (dsDNA). The interaction between TB and its aptamer led to the dissociation of dsDNA because TB has a higher affinity to TBA than the complementary strands. In the presence of exonuclease, aptamer was selectively digested and TB could be released for target recycling. Extended dsDNA was formed through HCR of the capture probe and two hairpin DNA strands (NH2-DNA1 and NH2-DNA1). Then, numerous europium multiwalled carbon nanotubes (Eu-MWCNTs) could be introduced through amidation reaction between NH2-terminated DNA strands and carboxyl groups on the Eu-MWCNTs, resulting in an increased ECL signal. The multiple amplification strategies, including the amplification of analyte recycling and HCR, and high ECL efficiency of Eu-MWCNTs lead to a wide linear range (1.0×10(-12)-5.0×10(-9) mol/L) and a low detection limit (0.23 pmol/L). The method was applied to serum sample analysis with satisfactory results.

Sandwich-format electrochemiluminescence assay for PDGF-BB using quantum dots–dendrimer nanocomposites as probe

This work describes a novel electrochemiluminescence aptasensor for highly sensitive detection of platelet-derived growth factor BB (PDGF-BB) using aptamer functionalized CdS quantum dots–polyamidoamine as probe (CdS QDs–PAMAM–Apt). CdS QDs–PAMAM nanocomposites were synthesized by one-pot synthesis in methanol. The prepared nanocomposites were linked with the NH2–aptamer 2 (Apt2) of PDGF-BB to form the CdS QDs–PAMAM–Apt2 probe by glutaraldehyde as coupling reagent. For constructing the aptasensor, MWCNTs–chitosan composites and NH2–aptamer 1 (Apt1) with the same base sequence as Apt2 were immobilized on the electrode by the self-assembled method to recognize the target protein PDGF-BB. In the presence of PDGF-BB, the structure of sandwiched format was formed between the Apt1 and the CdS QDs–PAMAM–Apt2 probe, thereby resulting in a proportional increase of ECL emission. Thanks to the efficient and stable ECL emission of CdS QDs–PAMAM dendrimer and the advantage of MWCNTs for accelerating the electron transfer, the highly sensitive detection of PDGF-BB with a detection limit of 0.13pM was achieved. The linear range is from 0.5pM to 1nM. The present protocol was applied to the analysis of PDGF-BB in human serum samples. The recoveries of PDGF-BB in human serum samples are 87.2–113% and RSD values are less than 3.6%.

Au nanoparticles decorated C60 nanoparticle-based label-free electrochemiluminesence aptasensor via a novel “on-off-on” switch system

Herein, a label-free electrochemiluminescence (ECL) aptasensor for highly sensitive determination of kanamycin was developed based on a novel “on-off-on” switch system. The first “switch on” state with remarkably high ECL intensity was obtained by the tri-layer composite films modified glassy carbon electrode towards the S2O82−-O2 system. To be specific, the first layer was the Au nanoparticles decorated C60 nanoparticles (abbreviation as Au@nano-C60) as inner-layer which was prepared by the in situ generating of Au nanoparticles onto the surface of bovine serum albumin decorated nano-C60 nanoparticles. Then poly-l-histidine was first selected as a novel coreactant of S2O82−-O2 system and it was adsorbed on the Au@nano-C60 modified electrode as inter-layer. Finally, a self-assembling layer of colloidal Au nanoparticles (AuNPs) was the outer-layer. The three layers were interaction by the Au–N covalent bond which could achieve a desirable initial amplified ECL signal. Successively, the capture probes (CPs) of the aptamer for the target of kanamycin was anchored on the resultant tri-layer composite films modified electrode. Later, the “switch off” state was made by the quenching effect of hemin/G-quadruplex DNAzymes towards S2O82−-O2 system according to the DNA hybridization reaction of an assistant probes (APs, guanine-rich nucleic acid) with CPs which could generate a large amount of hemin/G-quadruplex DNAzymes in the presence of hemin with a simple and label-free process. As expected, the second “switch on” state was the ECL signal recovery when the target of kanamycin was present, it is attributed to that the formation of the aptamer–kanamycin complex makes the quencher of hemin/G-quadruplex DNAzymes release from the sensing interface. With the association of “on-off-on” switch system, a linear response about 9 orders of magnitude for kanamycin detection was obtained from 0.15 nM to 170 mm as well as a detection limit down to 45 pM.

Signal-on electrochemiluminescent aptasensors based on target controlled permeable films.

A novel permeability gate-based electrochemiluminescent (ECL) aptasensor has been constructed by utilizing target-responsive polyelectrolyte-aptamer film deposited on the solid-state ECL electrode to control the rate of diffusion of a coreactant that triggers the ECL.

Ultrasensitive electrochemiluminescence aptasensor based on a graphene/polyaniline composite film modified electrode and CdS quantum dot coated platinum nanostructured networks as labels

A simple electrochemiluminescence (ECL) aptasensor for adenosine triphosphate (ATP) based on graphene/polyaniline (GR/PANI) composite films modified glassy carbon electrode (GCE) was successfully fabricated.

An amplified electrochemiluminescent aptasensor using Au nanoparticles capped by 3,4,9,10-perylene tetracarboxylic acid-thiosemicarbazide functionalized C60 nanocomposites as a signal enhancement tag

A novel electrochemiluminescent (ECL) signal tag of Au nanoparticles capped by 3,4,9,10-perylene tetracarboxylic acid-thiosemicarbazide functionalized C60 nanocomposites (AuNPs/TSC-PTC/C60NPs) was developed for thrombin (TB) aptasensor construction based on the peroxydisulfate/oxygen (S2O8(2-)/O2) system. For signal tag fabrication, the C60 nanoparticles (C60NPs) were prepared and then coated with 3,4,9,10-perylene tetracarboxylic acid (PTCA) by π-π stacking interactions. Afterwards, thiosemicarbazide (TSC) was linked with PTCA functionalized C60NPs via amidation for further assembling Au nanoparticles (AuNPs). Finally, detection aptamer of thrombin (TBA 2) was labeled on the ECL signal amplification tag of AuNPs/TSC-PTC/C60NPs. Herein, TSC, with the active groups of -NH2 and -SH, was selected and introduced into the ECL S2O8(2-)/O2 system for the first time, which could not only offer the active groups of -SH to absorb AuNPs for TBA 2 anchoring but also remarkably enhance the ECL signal of the S2O8(2-)/O2 system by the formation of TSC-PTC/C60NPs for signal amplification. Meanwhile, the sensing interface of a glassy carbon electrode (GCE) was modified by AuNPs/graphene (AuNPs/GR) nanocomposites with the large specific surface area and the active sites, followed by immobilization of thiol-terminated thrombin capture aptamer (TBA 1). With the formation of the sandwich-type structure of TBA 1, TB, and TBA 2 signal probes, a desirable enhanced ECL signal was measured in the testing buffer of an S2O8(2-)/O2 solution for detecting TB. The aptasensor exhibited a good linear relationship for TB detection in the range of 1 × 10(-5)-10 nM with a detection limit of 3.3 fM.

An efficient signal-on aptamer-based biosensor for adenosine triphosphate detection using graphene oxide both as an electrochemical and electrochemiluminescence signal indicator.

An efficient aptasensor was developed in which graphene oxide (GO) was employed as an indicator for both electrochemical impedance spectroscopy and electrochemiluminescence (ECL) signal generation. The aptasensor was fabricated by self-assembling the ECL probe of a thiolated adenosine triphosphate binding aptamer (ABA) tagged with a Ru complex (Ru(bpy)3(2+) derivatives) onto the surface of gold nanoparticle (AuNP) modified glassy carbon electrode (GCE). ABA immobilized onto AuNP modified GCE could strongly adsorb GO due to the strong π-π interaction between ABA and graphene oxide; ECL quenching of the Ru complex then takes place because of energy transfer and electron transfer, and a large increase of the electron transfer resistance (Ret) of the electrode. While in the presence of target adenosine triphosphate (ATP), the ABA prefers to form ABA-ATP bioaffinity complexes, which have weak affinity to graphene oxide and keep the graphene oxide away from the electrode surface, thus allowing the ECL signal enhancement, and in conjunction with the decrease of the Ret. Because of the high ECL quenching efficiency, unique structure, and electronic properties of graphene oxide, the Ret and ECL intensity versus the logarithm of ATP concentration was linear in the wide range from 10 pM to 10 nM with an ultra-low detection limit of 6.7 pM to 4.8 pM, respectively. The proposed aptasensor exhibited excellent reproducibility, stability, and outstanding selectivity, and ATP could be effectively distinguished from its analogues. More significantly, this efficient ECL aptasensor strategy based on GO acting both as an electrochemical and ECL signal indicator is general and can be easily extended to other biological binding events.

Biobar-Coded Gold Nanoparticles and DNAzyme-Based Dual Signal Amplification Strategy for Ultrasensitive Detection of Protein by Electrochemiluminescence

A dual signal amplification strategy for electrochemiluminescence (ECL) aptasensor was designed based on biobar-coded gold nanoparticles (Au NPs) and DNAzyme. CdSeTe@ZnS quantum dots (QDs) were chosen as the ECL signal probes. To verify the proposed ultrasensitive ECL aptasensor for biomolecules, we detected thrombin (Tb) as a proof-of-principle analyte. The hairpin DNA designed for the recognition of protein consists of two parts: the sequences of catalytical 8-17 DNAzyme and thrombin aptamer. Only in the presence of thrombin could the hairpin DNA be opened, followed by a recycling cleavage of excess substrates by catalytic core of the DNAzyme to induce the first-step amplification. One part of the fragments was captured to open the capture DNA modified on the Au electrode, which further connected with the prepared biobar-coded Au NPs-CdSeTe@ZnS QDs to get the final dual-amplified ECL signal. The limit of detection for Tb was 0.28 fM with excellent selectivity, and this proposed method possessed good performance in real sample analysis. This design introduces the new concept of dual-signal amplification by a biobar-coded system and DNAzyme recycling into ECL determination, and it is promising to be extended to provide a highly sensitive platform for various target biomolecules.

Application of bimetallic PtPd alloy decorated graphene in peroxydisulfate electrochemiluminescence aptasensor based on Ag dendrites decorated indium tin oxide device

In this paper, PtPd alloy decorated reduced graphene oxide (RGO/PtPd) was synthesized by a simple and green method. The RGO/PtPd could catalytic oxidation of H2O2 to form O2 and O2 acted as the coreactant of peroxydisulfate, could greatly amplify the ECL signal. Moreover, the RGO/PtPd exhibited higher catalytic activity compared to the RGO/Pt or RGO/Pd catalyst because of the synergetic effects of Pt and Pd and the enhanced electron transfer of RGO. Based on this, a highly sensitive and selective label-free electrochemiluminescence (ECL) aptasensor for detection of adenosine triphosphate was designed. In this process, RGO/PtPd could be regarded as mimicking enzyme, the effect of which was similar to horseradish peroxidase in generating O2 and also used as carriers to immobilize ssDNA2. With respect to sensitivity, Ag dendrites with hierarchical nanostructures were electrodeposited on working electrode of portable indium tin oxide device, which improved the electronic transfer rate as well as increased the amount of immobilized capture ssDNA1. Under the optimal conditions, the approach provided a linear range from 0.1pM to 50nM and a low detection limit of 0.037pM.

Ultrasensitive and Signal‐on Electrochemiluminescence Aptasensor Using the Multi‐tris(bipyridine)ruthenium(II)‐β‐cyclodextrin Complexes

AbstractAn ultrasensitive and signal‐on electrochemiluminescence (ECL) aptasensor to detect target protein (thrombin or lysozyme) was developed using the host‐guest recognition between a metallocyclodextrin complex and single‐stranded DNA (ss‐DNA). The aptasensor uses both the photoactive properties of the metallocyclodextrins named multi‐tris(bipyridine)ruthenium(II)‐β‐cyclodextrin complexes and their specific recognition with ss‐DNA, which amplified the ECL signal without luminophore labeling. After investigating the ECL performance of different multi‐tris(bipyridine)ruthenium(II)‐β‐cyclodextrin (multi‐Ru‐β‐CD) complexes, tris‐tris(bipyridine)‐ruthenium(II)‐β‐cyclodextrin (tris(bpyRu)‐β‐CD) was selected as a suitable host molecule to construct an atasensor. First, double‐stranded DNA (ds‐DNA) formed by hybridization of the aptamer and its target DNA was attached to a glassy carbon electrode via coupling interaction, which showed low ECL intensity with 2‐(dibutylamino) ethanol (DBAE) as coreactant, because of the weak recognition between ds‐DNA and tris(bpyRu)‐β‐CD. Upon addition of the corresponding protein, the ECL intensity increased when target ss‐DNA was released because of the higher stability of the aptamer‐protein complex than the aptamer‐DNA one. A linear relationship was observed in the range of 0.01 pmol/L to 100 pmol/L between ECL intensity and the logarithm of thrombin concentrations with a limited detection of 8.5 fmol/L (S/N=3). Meanwhile, the measured concentration of lysozyme was from 0.05 pmol/L to 500 pmol/L and the detection limit was 33 fmol/L (S/N=3). The investigations of proteins in human serum samples were also performed to demonstrate the validity of detection in real clinical samples. The simplicity, high sensitivity and specificity of this aptasensor show great promise for practical applications in protein monitoring and disease diagnosis.