Aptamer Sensor Research Articles

A label-free fluorometric aptasensor for adenosine triphosphate (ATP) detection based on aggregation-induced emission probe

Based on Aggregation-Induced Emission (AIE), the development of a label-free, simple and sensitive fluorometric aptasensor for adenosine triphosphate (ATP) detection is described. With ATP present, the aptamers will combine with ATP and the conformation of the aptamer will switch from a random coil to an antiparallel G-quadruplex, which impedes the digestion by exonuclease I (Exo I). Addition of 4,4 -(1E,1E)-2,2-(anthracene-9,10-diyl) bis (ethene-2,1-diyl) bis (N,N, N-trimethyl-benzenaminium iodide) (DSAI) into the solution will cause aggregation of DSAI on the surface of the aptamer/ATP complex and consequently give rise to strong emission. Additionally, a good linear relationship was observed under optimized conditions between the fluorescence intensities and the logarithm of ATP concentrations (R2 = 0.9908). The established aptamer sensor was highly sensitive and exhibited a low limit of detection of 32.8 nM, with superior specificity for ATP. It was also used in the quantification of ATP levels in human serum samples and demonstrated satisfactory recoveries in the scope of 93.2%–107.6%. The cellular ATP assay results indicated that the developed method can be used for monitoring ATP concentrations in cell extracts without the interference of other substances in the cells. This method offers several advantages such as simplicity, rapidity, low cost and excellent selectivity, which make it hold great potential for the detection of ATP in bioanalytical and biological studies.

New Strategy for Ultrasensitive Aptasensor Fabrication: D-A-D Constitution as a Charge Transfer Platform and Recognition Element.

Over the past decade, various sensing systems based on aptamers have attracted a great deal of studies directed at designing highly selective biosensors. In this paper, we described a new-style electrochemical aptamer sensor (aptasensor) via a donor-acceptor link substrate, which was characterized by electrochemical methods and other helpful characterization instruments. Molecules with D-A-D configuration always undergo an intrinsic signal amplification due to the elongation of the π-electron conjugation. Triphenylamine, a peripheral electron donor, has excellent hole-transport property and is able to assemble on the surface of glassy carbon electrode by π-π stacking interaction. To further improve the performance of the adenosine triphosphate (ATP) sensor, we chose diphenylfumaronitrile-containing electron-withdrawing group as the central core to promote charge transfer, which can also efficiently combine with aptamers by multihydrogen bond function. Surprisingly, the sensing platform showed a wide liner range from 0.1 pM to 100 nM, with a detection limit of 0.018 pM. We examined the ATP in human serum sample, indicating that the novel aptasensor based on D-A-D conjugated polymer holds great possibility for practical detection of ATP. Moreover, it is foreseeable that the conjugated polymers of the D-A structure will have promising application in the preparation of biosensors.

A comparison of the performance of voltammetric aptasensors for glycated haemoglobin on different carbon nanomaterials-modified screen printed electrodes

The integration of carbon nanomaterials into electrochemical aptasensors has gained significant interest in the recent years because of their high electrical conductivity, mechanical strength, and large surface area. However, no comparative study has been reported so far between different carbon nanomaterials for aptasensing applications. Here, we report, a comparative investigation of six carbon electrode materials (carbon, graphene (G), graphene oxide (GO), multi-wall carbon nanotube (MWCNT), single walled carbon nanotube (SWCNT) and carbon nanofiber (CNF)) on the performance of glycated haemoglobin (HbA1c) aptasensor prepared by physical adsorption. The aptamers were non-covalently immobilized on the six nanomaterial electrodes via π-π stacking interactions between the DNA nucleobases and the surface of the carbon material which creates a barrier to the electron transfer. However, upon binding of the target protein to the aptamer, the aptamer dissociates from the surface leading to enhancement of the electron transfer which represent the basis of the detection. The aptamer adsorption, sensors responses and selectivity of the different nanomaterials were compared showing better performance of the SWCNT-based sensor. The voltammetric SWCNT aptasensors showed high sensitivity and selectivity with detection limits of 0.13 pg/mL and 0.03 pg/mL for total haemoglobin (tHb) and HbA1c, respectively. The aptasensor showed selectivity against other proteins in the blood including cystic fibrosis transmembrane conductance regulator (CFTR), survival motor neuron (SMN), dedicator of cytokinesis 8 (DOCK8), signal transducer and activator of transcription 3 (STAT3). This SWCNT aptasensor was superior to the reported detection assays for HbA1c in terms of sensitivity, selectivity and cost. Moreover, our results demonstrate that the choice of the carbon nanomaterial can have a profound impact on the biosensing performance.

Optimization of fluorescent aptamer sensor for ATP detection

Abnormal concentrations of ATP are associated with many diseases and cancers, and quantitative detection of ATP is thus of great importance for disease diagnosis and prognosis. In order to obtain a simple and quickly detection method of ATP by fluorescence aptasensor, the principle of ATP detection based on resonance energy transfer between FAM and GO was studied. And then the optimization of process conditions was carefully researched. The results showed that when the concentration and reaction time of GO was 10μ g/mL and 5min, respectively, the quenching light intensity was the easiest to recover. With the optimized parameters, the concentration of ATP in the range of 1-250 μ mol/L had a good linear relationship with the fluorescence intensity, and the detection limit was 1μ mol/L. Besides, the selectivity experiments proved the selectivity of the constructed aptasensor toward ATP. This method creates the opportunity to detect a broad range of targets by using appropriate aptamers, holding great potential in the field of food safety and biomedical diagnostics.

An aptasensor for troponin I based on the aggregation-induced electrochemiluminescence of nanoparticles prepared from a cyclometallated iridium(III) complex and poly(4-vinylpyridine-co-styrene) deposited on nitrogen-doped graphene.

An ultrasensitive electrochemiluminescence (ECL) disposable aptamer sensor (aptasensor) is presented for detection of myocardial infarction biomarker by quantification of troponin I in blood serum. A screen-printed electrode was modified with (a) aptamer-modified gold nanoparticles, (b) cyclometallated iridium(III)-poly-4-vinylpyridine nanoparticles, and (c) nitrogen-doped graphene in order to increase the loading capacity and conductivity of the aptasensor. If the aptasensor is exposed to troponin I, it will bind to the aptamer and desorb the aptamer from gold nanoparticles and the surface of the electrode. This generates an enhancement in ECL emission depending on troponin I concentration. ECL emission is strongly improved by aggregation-induced phenomenon, which is caused by inhibition of the water and oxygen quenching effect on the iridium complex ECL in aqueous media. Under optimum conditions, the aptasensor has a wide dynamic range that extends from 0.1 pM to 10nM, with a 20 fM detection limit (S/N = 3) and a relative standard deviation of 3.1%. The ECL aptasensor was successfully applied to 20 individual human serum for the detection of troponin I biomarker. Graphical abstract Schematic presentation of electrochemiluminescence aptamer assay fabrication for detection of Troponin I. Carbon screen printed electrode (CSPE) was modified with nitrogen doped graphene (NG), gold nanoparticles (AuNPs), cyclometallated iridium(III)-polyvinylpyridine polymer nanoparticles, ionic liquid and bovine serum albumin.

Liquid crystal-based aptamer sensor for sensitive detection of bisphenol A

Bisphenol A (BPA) is an environmental poison that seriously affects the health of humans. Herein, a novel, label-free, time-saving liquid crystal-based aptamer sensor for the detection of BPA was developed based on the specific binding of an aptamer and its target. In this study, the surface of the sensor was assembled with (3-aminopropyl)triethoxysilane & dimethyloctadecyl[3-(trimethoxysilyl) propyl]ammonium chloride and the amino-aptamer was attached to this film via glutaradehyde links. Through a specific reaction, BPA reacts with the BPA aptamer forming an ‘aptamer–BPA’ complex that changes the conformation of the BPA aptamer. Through spatial size effects, the large size of the complex disturbs the nematic orientation of 4-cyano-4′-pentybiphenyl in the film on this surface from homeotropic to random, resulting in an image that changes from ‘dark’ to ‘bright’ under a polarized light microscope. The limit of detection of this system is 600 pM. Atomic force microscopy was also employed to further validate the results.

Simple and Label‐Free Liquid Crystal‐based Optical Sensor for Highly Sensitive and Selective Endotoxin Detection by Aptamer Binding and Separation

AbstractLipopolysaccharides (LPS), also commonly known as lipoglycans and endotoxins, can often accidentally contaminate recombinant protein therapeutics and bacteria‐derived plasmid DNA vaccines. Since LPS can induce fever, hypotension, shock, and even death, their early and sensitive detection is necessary in relevant bioassays and for parenteral drug administration and/or biotherapeutics. In this study, an optical biosensor was developed using LPS‐specific single‐stranded DNA aptamers as LPS‐selective probes. (3‐Aminopropyl) triethoxysilane and glutaraldehyde were used as linkers to immobilize the LPS high‐affinity aptamer on glass. Subsequently, each modification step was characterized by the optical responses obtained from liquid crystals using a polarized optical microscope. The sensor's ability to detect LPS was confirmed using a broad LPS detection range (5.5 pg/mL ‐ 100 ng/mL). Despite the presence of plasmid DNA, RNA, and bovine serum albumin, the aptamer sensor showed high selectivity for LPS and could be regenerated for reuse at a low pH, thus, providing a promising option for detecting LPS in complex, low pH environments.

Split Dapoxyl Aptamer for Sequence-Selective Analysis of Nucleic Acid Sequence Based Amplification Amplicons.

Hybridization probes have been used for the detection of single nucleotide variations (SNV) in DNA and RNA sequences in the mix-and-read formats. Among the most conventional are Taqman probes, which require expensive quantitative polymerase chain reaction (qPCR) instruments with melting capabilities. More affordable isothermal amplification format requires hybridization probes that can selectively detect SNVs isothermally. Here we designed a split DNA aptamer (SDA) hybridization probe based on a recently reported DNA sequence that binds a dapoxyl dye and increases its fluorescence ( Kato, T.; Shimada, I.; Kimura, R.; Hyuga, M., Light-up fluorophore-DNA aptamer pair for label-free turn-on aptamer sensors. Chem. Commun. 2016 , 52 , 4041 - 4044 ). SDA uses two DNA strands that have low affinity to the dapoxyl dye unless hybridized to abutting positions at a specific analyte and form a dye-binding site, which is accompanied by up to a 120-fold increase in fluorescence. SDA differentiates SNV in the inhA gene of Mycobacterium tuberculosis at ambient temperatures and detects a conserved region of the Zika virus after isothermal nucleic acid sequence based amplification (NASBA) reaction. The approach reported here can be used for detection of isothermal amplification products in the mix-and-read format as an alternative to qPCR.

Dual-signal aptamer sensor based on polydopamine-gold nanoparticles and exonuclease I for ultrasensitive malathion detection

A highly sensitive dual-signal aptamer sensor based on polydopamine-gold nanoparticles (PDA-AuNPs) and exonuclease I (Exo I) was developed for detection of malathion. Compared with traditional sensing elements, aptamer has many advantages, such as high affinity, superior specificity, strong stability and easy modification. The electrodeposition synthesis of PDA-AuNPs gave excellent biocompatibility and electrical conductivity on the sensor. With the addition of malathion, the specific interaction between malathion and its aptamer forced the aptamer to detach from the electrode surface and induced the capture probe to form a hairpin structure on the electrode surface. Exo I was added to motivate the autocatalytic target cycling which remarkably increased the current change of electrochemical signal over 2 times. Therefore, the promising strategy gave rise to an optional dual-signal current readout in both the signal-on of Fc and the signal-off of Tn. In this work, the prepared biosensor exhibited high sensitivity to malathion via the combination of dual-signal design and autocatalytic target cycling amplification. Under the optimized conditions, the proposed sensor showed a wide linear range from 0.5 to 600 ng/L malathion. It also exhibited excellent specificity, acceptable repeatability and good stability. The application of real samples obtained satisfactory recovery results, demonstrating a promising potential in food safety analysis.

Monitoring 2,3',5,5'-tetrachlorobiphenyl with a rapid and sensitive environmental aptamer sensor.

Polychlorinated biphenyl (PCB) detection in the environment is significant for both environmental protection and human health. Herein, a highly sensitive aptamer sensor has been established by employing a 2,3',5,5'-tetrachlorobiphenyl (PCB72) targeting aptamer as a highly specific recognition element and a gold/silver (Au@Ag) nanocomposite as the surface-enhanced Raman spectroscopy (SERS) substrate for detecting environmental PCB72. The Au@Ag nanoparticles (NPs) exhibit a strong SERS enhancement and provide an efficient substrate for immobilizing the PCB72 aptamer and Raman signal labelled molecule, 4-mercaptobenzoic acid (4-MBA). The targeted PCB72 could competitively bind with the PCB72 aptamer, resulting in a few aptamers sticking to the Au@Ag NPs and the "hot spot" strengthening effect of the substrate. Under optimal conditions, this aptamer sensor exhibits great performance with high sensitivity, excellent selectivity and stability for the monitoring of PCB72, which shows an excellent linear correlation ranging from 1 to 1000 pg mL-1 with a limit of detection of 0.3 pg mL-1. Furthermore, this aptamer assay exhibits high specificity and selectivity for PCB72 with the detection error of less than 0.27 for other PCBs and 0.21 for other interfering species, even if the coexisting interferents are 100-fold concentration in the system. Additionally, the recognition mechanism of the binding of aptamers to PCB72 is analyzed via UV-vis spectroscopy and molecular docking simulations, which suggest that PCB72 could insert into the aptamers. Furthermore, this method is successfully utilized for PCB72 detection in real water samples with a simple pre-treatment. In general, this work provides a new and effective method using an environmental aptamer sensor for rapid and sensitive PCB72 detection.

Thermodynamic investigation of kissing-loop interactions

Kissing loop interactions (KLIs) are a common motif that is critical in retroviral dimerization, viroid replication, mRNA, and riboswitches. In addition, KLIs are currently used in a variety of biotechnology applications, such as in aptamer sensors, RNA scaffolds and to stabilize vaccines for therapeutics. Here we describe the thermodynamics of a basic intramolecular DNA capable of engaging in a KLI, consisting of two hairpins connected by a flexible linker. Each hairpin loop has a five-nucleotide complementary sequence theoretically capable of engaging in a KLI. On either side of each loop is two thymines which will not engage in kissing but are present to provide more flexibility and optimal KLI positioning. Our results suggest that the KLI occurs even at physiological salt levels, and that the KLI does not alter the thermodynamics and stability of the two stem structures. The KLI does not involve all five nucleotides, or at least each base-pair stack is not making full contact. Adding a second strand complementary to the bottom of the kissing complex removes flexibility and causes destabilization of the stems. The KLI of this less flexible complex is maintained but the TM is reduced, indicating an entopic penalty to its formation.

Amplified Split Aptamer Sensor Delivered Using Block Copolymer Nanoparticles for Small Molecule Imaging in Living Cells.

We develop a novel amplified split aptamer sensor for highly sensitive detection and imaging of small molecules in living cells by using cationic block copolymer nanoparticles (BCNs) with entrapped fluorescent conjugated polymer as a delivery agent. The design of a split aptamer as the initiator of hybridization chain reaction (HCR) affords the possibility of enhancing the signal-to-background ratio and thus allows high-contrast imaging for small molecules with relatively weak interactions with their aptamers. The novel design of using fluorescent cationic BCNs as the nanocarrier enables efficient and self-tracking transfection of DNA probes. Results reveal that BCNs exhibit high fluorescence brightness allowing direct tracking of the delivery location. The developed amplified split aptamer sensor is shown to have high sensitivity and selectivity for in vitro quantitative detection of adenosine triphosphate (ATP) with a detection limit of 30 nM. Live cell studies show that the sensor provides a "signal on" approach for specific, high-contrast imaging of ATP. The DNA sensor based HCR system may provide a new generally applicable platform for detection and imaging of low-abundance biomarkers.

A label-free impedimetric aptasensor for the detection of Bacillus anthracis spore simulant

Herein, we report an impedimetric DNA-based aptamer sensor for a single-step detection of B. anthracis spore simulant (B. cereus spore). Specifically, we designed a miniaturized label-free aptasensor for B. cereus spores based on a gold screen-printed electrode functionalized with B. cereus spores-binding aptamer (BAS-6R). Several parameters were optimized to fabricate the aptasensor such as the concentration of DNA aptamer solution (0.5 µM), the time (48 h), the temperature (4 °C), and the pH (7.5) for aptamer immobilization on the working electrode surface. Once the aptasensor was developed, it was tested against B. cereus spores 14579 evaluating the effect of incubation time and MgCl2 concentration. Under the optimized conditions (incubation time equal to 3 h and absence of MgCl2), B. cereus spores 14579 were detected with a linear range between 104 CFU/ml and 5 × 106 CFU/ml and a detection limit of 3 × 103 CFU/ml. Furthermore, the study of selectivity toward B. cereus 11778, B. subtilis, Legionella pneumophila, and Salmonella Typhimurium has demonstrated the capability of this sensor to detect B. cereus spores, proving the suitability of the DNA-based sensing element combined with a portable instrument for a label-free measurement on site of B. anthracis spore simulant.

Double strand DNA functionalized Au@Ag Nps for ultrasensitive detection of 17β-estradiol using surface-enhanced raman spectroscopy

A detection method for 17β-estradiol (E2) using surface-enhanced Raman scattering (SERS)-based aptamer sensor was presented. Raman reporter molecule Cy3 labeled E2-aptamer and DNA functionalized gold-silver core-shell nanoparticles (Au@Ag CS NPs) offered SERS with high sensitivity and selectivity. Based on the fabricated double strand DNA-immobilized gold-silver core-shell nanoparticles (Au@Ag NPs), SERS signal intensity of Raman reporter changed with the number of Cy3-labeled aptamer attached to the core-shell nanoparticles due to the strong binding affinity between the aptamers and E2 with different concentrations. A wide linear range from 1.0 × 10–13 to 1.0 × 10−9 was obtained for the detection of E2, with a low detection limit of 2.75 fM. This proposed method showed highly sensitive and selective for detecting E2, and could be used to determine E2 in actual samples.

Facile combination of beta-cyclodextrin host-guest recognition with exonuclease-assistant signal amplification for sensitive electrochemical assay of ochratoxin A

Smartly coupling exonuclease-induced target recycling signal amplifications with β-cyclodextrin host-guest recognition, a novel “signal-on” aptamer sensor for sensitive determination of ochratoxin A (OTA) was proposed for the first time. Firstly, the formation of double-strand DNA (dsDNA) was occurred by hybridizing OTA aptamer with its complementary DNA (cDNA) and as the probe DNA the cDNA at its 3′ terminal was labeled with methylene blue (MB). Next, when OTA was present, the aptamer tended to form aptamer-OTA complex with conformation of G-quadruplex instead of aptamer-cDNA duplex, leading to thus the probe DNA separating from dsDNA complex. Then the RecJf exonuclease was added, demolishing partially G-quadruplex structure and releasing a certain number of OTA. Sequentially, those released OTA would continue to react with the rest of aptamer in dsDNA, drawn into development of a new round of G-quadruplex complex, where the target cycling was realized. Meanwhile, as a signal molecule, MB modified on cDNA was liberated along with the cDNA being digested into monoucleotides by RecJf exonuclease, capable of diffusing onto the electrode surface due to host-guest recognition with β-cyclodextrin, whereupon the signal was enriched and yielded. In this way, cycles of target with continuous output of signal indicators were undergone, in which the detection of target was in return fulfilled with signal amplification owing to the joint endeavor of exonuclease and β-cyclodextrin. Under the optimal conditions, the raising signal maintained a linear relation with the logarithm of the target concentrations ranging from 10 pg/mL to 10.0 ng/mL and the detection limit reached as low as 3 pg/mL. This brand-new strategy was simple and low-cost but satisfactory in terms of detection limit, range and sensitivity, in all possibility to be applied extensively for diverse targets detection by easily alternating the corresponding aptamers.

Bimetallic cerium/copper organic framework-derived cerium and copper oxides embedded by mesoporous carbon: Label-free aptasensor for ultrasensitive tobramycin detection

We reported a novel bimetallic cerium/copper-based metal organic framework (Ce/Cu-MOF) and its derivatives pyrolyzed at different temperatures, followed by exploiting them as the scaffold of electrochemical aptamer sensors for extremely sensitive detection of trace tobramycin (TOB) in human serum and milk. After the calcination at high temperature, the meal coordination centers (Ce and Cu) were transferred to metal oxides containing various chemical valences, such as Ce(III), Ce(IV), Cu(II) and Cu(0), which were embedded within the mesoporous carbon network originated from the organic ligands (represented by CeO2/CuOx@mC). Owning to the strong synergistic effect among the metal oxides, mesoporous carbon, and small cavities and open channels of MOF, the as-prepared CeO2/CuOx@mC nanocomposites not only possess good electrochemical activity but also exhibit strong bioaffinity toward the aptamer strands. By comparing the electrochemical biosensing peroformances using on the Ce/Cu-MOF- and the series of CeO2/CuOx@mC-based aptasensors, the constructed CeO2/CuOx@mC900-based (calcinated at 900 °C) aptasensor exhibits an extremely low detection limit of 2.0 fg mL−1 within a broad linear TOB concentration range from 0.01 pg mL−1 to 10 ng mg L−1. It demonstrates that the proposed aptasensor is substantially superior to those previously reported in the literature, along with high selectivity, good stability and reproducibility, and acceptable applicability in human serum and milk. Thereby, the newly fabricated aptasensing approach based on bimetallic CeO2/CuOx@mC has a considerable potential for the quantitative detection of antibiotics in the food safety and biomedical field.

Graphene nanocomposites modified electrochemical aptamer sensor for rapid and highly sensitive detection of prostate specific antigen

Prostate specific antigen (PSA) is a widely used marker for the diagnosis of prostate cancer, and the increasing attention has been attracted on the development of rapid assay using biosensing technology. However, it remains challenging for the sensitive and selective detection of PSA in clinical samples. Here, we report a label-free microfluidic paper-based analytical device for highly sensitive electrochemical detection of PSA. The paper device was fabricated with wax printing to generate hydrophobic and hydrophilic layers for the construction of microfluidic channel, followed by screen-printing of three electrodes including working, counter and reference electrode. Gold nanoparticles (AuNPs)/reduced graphene oxide (rGO)/thionine (THI) nano composites were synthesized and characterized, which were coated onto working electrodes for the immobilization of DNA aptamer probe. THI servers as the electrochemical mediator to transduce the biological recognition between DNA aptamer and PSA, and the excellent conductivity of AuNPs and rGO also play a significant role of electron transfer, leading to a sensitive detection for PSA, able to detect PSA as low as 10 pg mL−1, with a linear range from 0.05 to 200 ng mL−1. We demonstrated that our electrochemical sensor for the detection of clinical serum samples, indicating that our sensor would provide a new platform for low cost, sensitive and point-of-care diagnosis of prostate cancer.

A novel ratiometric electrochemical assay for ochratoxin A coupling Au nanoparticles decorated MoS2 nanosheets with aptamer

A novel aptamer sensor for ratiometric ochratoxin A (OTA) detection was constructed based on the host-guest competition mode. Firstly, the aptamer modified with methylene blue (MB) at its terminal was fixed on the surface of the gold electrode via the host-guest recognition with β-cyclodextrin (β-CD). Then the MoS2 nanosheets/Au NPs nanocomposites were used for the signal amplification. When OTA was present, the configuration of MB-aptamer transformed from vertical linear to G-quadruplex, thus making itself unable to get close to the electrode surface and causing a decrease in the redox current of the MB-aptamer. Whereas the free ferrocenecarboxylic acid could recognize and occupy the vacant site and produce signals in the current, resulting in the “ratiometric” effect. Consequently, on the basis of the “dual-signal” principle, a highly sensitive detection of OTA was made, indicating that the OTA concentration had a linear response to the signal of sensor between 0.1 nM and 50 nM, and as low as 0.06 nM OTA had been detected. Overall, this method for OTA detection was constructed easily with high sensitivity and specificity, providing a significant platform for highly efficient detection of OTA in biological samples.

Harnessing the affinity of magnetic nanoparticles toward dye-labeled DNA and developing it as an universal aptasensor revealed by lipopolysaccharide detection

In current study, we have found that several magnetic nanoparticles (MNPs) are able to absorb DNA molecules, and surface engineering would be beneficial to tune such interaction. We then have focused on the assembly of polyethylenimine (PEI) coated MNPs (PEI-MNPs) with ssDNA (single-stranded DNA) and found this assembly is mediated by two forces, namely the electrostatic interactions of surface charges of MNPs and the phosphate backbones of DNA; as well as the coordination of exterior iron ions (especially Fe3+) of MNPs and DNA phosphate backbones. The fluorescence of dye-labeled DNA is significantly quenched when being complexed with PEI-MNPs, which is proved to be caused by static quenching. This PEI-MNPs interact with DNA, which could be harnessed for devising a novel type of aptasensor. This has been examplified by the selective and sensitive detection of lipopolysaccharide (LPS). The LOD (limit of detection) is ∼35 ng/mL and the linear range from 50 ng/mL to 10 μg/mL. Compared with widely used graphene oxide (GO)‒ssDNA aptamer sensors, we also have demonstrated that the PEI-MNPs based sensor is able to better avoid non-specific DNA displacement by interfering proteins, generating more satisfactory signal-to-background ratio. Our proposed sensor could be a supplement to classic GO‒DNA sensors. In summary, our work provides fundamental understanding of MNPs‒DNA interactions and also paves the way for developing novel MNPs based sensing approaches, which would contribute to nano‒bio interface and DNA-assisted bio-analysis, DNA-coordinated nano-materials and DNA-directed assembly.

Rational engineering of synergically stabilized aptamer-cDNA duplex probes for strand displacement based electrochemical sensors

Electrochemical aptamer (EA) sensors employing electrode-bound aptamer-cDNA (cDNA: complementary DNA) duplex probes are simple and versatile sensing platforms for detection of many analytes. Target recognition in these sensors is a strand displacement process where target binding to aptamer displaces the cDNA strand. Despite many displacement-type EA sensors, a common drawback exists that sensor response rate is relatively slow (typically with target binding equilibrium time ranging from 0.5 to 3 h), which is mainly caused by a suboptimal conformation of the aptamer sequence in the conventional aptamer-cDNA duplex probe containing a relatively large number of aptamer-involved base pairs. Realizing that the key to optimizing sensor sensitivity and response rate of displacement-type EA sensors lies in engineering of a class of aptamer-cDNA probes with moderate duplex stability and a minimum amount of aptamer-involved base pairs, we explore here a unique means for rational design of this type of aptamer-cDNA duplex probes by using the concepts of synergic stabilization and base-pair regulation. We have integrated a short non-aptamer base-pair sequence and a short aptamer-involved base-pair sequence to engineer a class of synergically stabilized aptamer-cDNA duplex probe. Through further regulation of the number of aptamer-involved base pairs within the duplex we can tune not only the signal increase but also the response rate of these synergically stabilized duplex probes to achieve optimal sensor performance. The rationally engineered synergically stabilized aptamer-cDNA duplex probes are sensitive and exhibit significantly enhanced response rates (with target binding equilibrium times of 1 min for ATP and 2 min for cocaine). The concepts of synergic stabilization and base-pair regulation can be generalized to any aptamer-cDNA duplex probes and will help guide further development of displacement-type EA sensors for sensitive and quick detection of a wide variety of target analytes.