Effects of citric acid on antioxidant system and carbon-nitrogen metabolism of Elymus dahuricus under Cd stress.

Exogenous citric acid (CA), which acts as an important intermediate product of the tricarboxylic acid (TCA) cycle, can enhance the TCA cycle activity and activate the branched operation of the TCA cycle, thus providing energy required for resistance to adverse conditions. However, the effects of CA application on TCA cycle-related metabolism under cadmium (Cd) were less reported. To investigate the effects of CA on the Cd tolerance of Dahurian wildrye grass (Elymus dahuricus), the growth, Cd accumulation, antioxidant systems and metabolic pathways of leaves and roots were investigated by a potted soil experiment with Cd (50 mg/kg) and CA (4 mmol/L) treatments. The results showed that Cd stress seriously affected growth and induced the production of reactive oxygen in clover leaves and roots, leading to membrane peroxidation and activation of the antioxidant defense system. Exogenous CA could not only effectively relieve the inhibition of Cd stress on growth and reduce the amount of reactive oxygen by increasing the antioxidant capacities but could also promote an increase in root Cd content. Metabolomic results showed that the application of CA increased the contents of sugars, sugar alcohols, and resistant substances, and promoted the metabolism of amino acids including γ-aminobutyric acid (GABA). These alterations contributed the significant enhancement of the Cd resistance, which may be related to the changes in the TCA cycle activity and the metabolism of the shikimic acid pathway in leaves and roots as well as GABA shunt in roots.

Novel 2D-DNA-Nanoprobe-Mediated Enzyme-Free-Target-Recycling Amplification for the Ultrasensitive Electrochemical Detection of MicroRNA.

In this work, on the basis of a new 2D DNA nanoprobe (DNP) and an enzyme-free-target-recycling amplification, an electrochemical biosensor is developed for the ultrasensitive detection of microRNA-21 (miRNA-21). Herein, two ferrocene-labeled bipedal DNPs, which show small steric hindrance and strong stability, are prepared on the basis of the mechanism of the proximity-ligation assay (PLA), improving the space utilization. In the presence of the target, miRNA-21, and a hairpin DNA strand, the DNP will collapse, and then two ferrocene-labeled DNA strands and the miRNA-21 will be simultaneously released from the electrode surface through toehold-mediated strand-displacement reactions (TSDRs), leading to a decrease in the electrochemical signal and realization of enzyme-free target recycling. As a result, the one input target, miRNA-21, could release 2 N ferrocene-labeled DNA strands, achieving a dramatic decrease in the electrochemical signal. Combining DNPs and enzyme-free target recycling, this proposed biosensor showed a linear dependence with miRNA-21 concentration, ranging from 1.0 fM to 10 nM with a detection limit of 0.31 fM. In addition, it is worth mentioning that this biosensor can be regenerated through incubating with three assistant-DNA strands, realizing the reuse of raw materials. Surprisingly, the elaborated biosensor provides a novel strategy for building controllable DNA nanoprobes for the sensitive detection of various biomarkers.

A Sensitive Electrochemical Aptasensor for Thrombin Detection Based on Electroactive Co-Based Metal-Organic Frameworks with Target-Triggering NESA Strategy.

In this work, an improved target-triggering nicking enzyme signaling amplification (NESA) strategy as signal enhancer has been fabricated to obtain a sensitive electrochemical thrombin (TB) biosensor combined with PtPd NPs decorated electroactive Co-based metal-organic frameworks (Co-MOFs/PtPdNPs) as a redox mediator. Traditionally, in the NESA strategy, only one of the output double strands DNA is available in the next cycle. However, in this work, all of the output DNA involved in the improved NESA strategies could be further employed, resulting in high utilization of output DNA, which further enhanced signal amplification and sensitivity of the biosensor. In addition, the electroactive Co-MOFs were not only used as nanocarriers but also acted as signal labels, avoiding adding extra redox media. Simultaneously, in the presence of H2O2, PtPd NPs decorated on the Co-MOFs act the same as horseradish peroxidase to promote the oxidation of H2O2, further promoting the conversion of Co2+ to Co3+, leading to electrochemical signal amplification. With such design, the TB biosensor exhibited good sensitivity from 1 pM to 30 nM with a detection limit of 0.32 pM. This new NESA strategy with high utilization of output DNA can supply one efficient approach to improve signal amplification, which also open an avenue for sensitivity enhancement in detection of analytes.

Cu/Mn Double-Doped CeO2 Nanocomposites as Signal Tags and Signal Amplifiers for Sensitive Electrochemical Detection of Procalcitonin.

Nanomaterials themselves as redox probes and nanocatalysts have many advantages for electrochemical biosensors. However, most nanomaterials with excellent catalytic activity cannot be directly used as redox probe to construct electrochemical biosensor because the redox signal of these nanomaterials can only be obtained in strong acid or alkali solution at high positive or negative potential, which greatly limits their applications in biologic assay. In this study, Cu/Mn double-doped CeO2 nanocomposite (CuMn-CeO2) was synthesized to use as signal tags and signal amplifiers for the construction of electrochemical immunosensor for sensitive assay of procalcitonin (PCT). Herein, CuMn-CeO2 not only possesses excellent catalytic activity toward H2O2 for signal amplification, but also can be directly used as redox probe for electrochemical signal readout achieved in neutral mild buffer solution at low positive potential. Importantly, since doping Cu, Mn into CeO2 lattice structure can generate extra oxygen vacancies, the redox and catalytic performance of obtained CuMn-CeO2 was much better than that of pure CeO2, which improves the performance of proposed immunosensor. Furthermore, CuMn-CeO2 can be implemented as a matrix for immobilizing amounts of secondary antibody anti-PCT by forming ester-like bridging between carboxylic groups of Ab2 and CeO2 without extra chemical modifications, which greatly simplifies the preparative steps. The prepared immunosensor exhibited a wide linear range of 0.1 pg mL-1 to 36.0 ng mL-1 with a low detection limit of 0.03 pg mL-1. This study implements nanomaterial themselves as redox probes and signal amplifiers and paves a new way for constructing electrochemical immunosensor.

Highly Effective Protein Converting Strategy for Ultrasensitive Electrochemical Assay of Cystatin C.

In this work, a highly effective protein converting strategy based on immunoreaction-induced DNA strand displacement and T7 Exonuclease (T7 Exo)-assisted protein cyclic enzymatic amplification for ultrasensitive detection of cystatin C was described. Herein, Au@Fe3O4 as magnetic separator was labeled by antibody 1-conjungated DNA (DNA1) and the DNA substrate of T7 Exo (DNA3) which initially hybridized with output DNA (S1) to form a stable S1/DNA1 duplex (S1/DNA3). Antibody 2 was labeled by competing DNA (DNA 2). In the presence of cystatin C, sandwich immunoreaction would induce proximity hybridization between DNA2 and DNA3 and thus displace S1 from the S1/DNA3 duplex with formation of a stable DNA2/DNA3 duplex, realizing the conversion of input target cystatin C into output S1. To enhance the conversion ratio, the DNA2/DNA3 duplex was then digested by T7 Exo with release of DNA2 which could act as competing DNA again to displace S1 from the S1/DNA3 duplex in adjacent locations and initiate another cleavage reaction. Through such a cyclic process, each input cystatin C could induce more than one output S1, enhancing detection sensitivity. A hairpin DNA modified electrode was used to capture the output S1, and then, a hybridization chain reaction is triggered on the biosensor surface. Then, thionine as electron mediator was embedded into the dsDNA polymers to produce a detection signal. The electrochemical biosensor exhibited a much wider linear range of 0.01 pg mL(-1) to 30 ng mL(-1) with low detection limit of 3 fg mL(-1). Moreover, this method introduced protein unrelated to nucleic acids into the realm of potential inputs for translation, which might create a new immunoassay method for sensitive detection of protein.

Film-forming, stable, conductive composites of polyhistidine/graphene oxide for electrochemical quantification of trace Pb2.

Nanomaterials with unique properties, such as good film-formation and plentiful active atoms, play a vital role in the construction of electrochemical sensors. In this work, an in situ electrochemical synthesis of conductive polyhistidine (PHIS)/graphene oxide (GO) composite film (PHIS/GO) was designed to construct an electrochemical sensor for the sensitive detection of Pb2+. Herein, GO as an active material can directly form homogeneous and stable thin films on the electrode surface because of its excellent film-forming property. Then GO film was further functionalized by in situ electrochemical polymerization of histidine to obtain plentiful active atoms (N). Due to strong van der Waals forces between GO and PHIS, PHIS/GO film exhibited high stability. Furthermore, the electrical conductivity of PHIS/GO films was greatly improved by in situ electrochemical reduction technology and the plentiful active atoms (N) in PHIS are profitable for adsorbing Pb2+ from solution, tremendously enhancing the assay sensitivity. With the above unique property, the proposed electrochemical sensor showed high stability, a low detection limit (0.045 µg L-1) and a wide linear range (0.1-300 µg L-1) for the quantification of Pb2+. The method can also be extended to the synthesis of other film-forming nanomaterials to functionalize themselves and widen their potential applications, avoiding the addition of non-conductive film-forming substances.

Programmable High-Speed and Hyper-Efficiency DNA Signal Magnifier.

Herein, a programmable dual-catalyst hairpin assembly (DCHA) for realizing the synchronous recycle of two catalysts is developed, displaying high reaction rate and outstanding conversion efficiency beyond traditional nucleic acid signal amplifications (NASA). Once catalyst I interacts with the catalyst II, the DCHA can be triggered to realize the simultaneous recycle of catalysts I and II to keep the highly concentrated intermediate product duplex I-II instead of the steadily decreased one in typical NASA, which can accomplish in about only 16 min and achieves the outstanding conversion efficiency up to 4.54 × 108 , easily conquering the main predicaments of NASA: time-consuming and low-efficiency. As a proof of the concept, the proposed DCHA as a high-speed and hyper-efficiency DNA signal magnifier is successfully applied in the rapid and ultrasensitive detection of miRNA-21 in cancer cell lysates, which exploits the new generation of universal strategy for the applications in biosensing assay, clinic diagnose, and DNA nanobiotechnology.

Facile fabrication and low-temperature bonding of Cu@Sn-Bi core-shell particles for conductive pastes.

The rapid development of flexible wearable electronics arouses huge demand for low-temperature sintering metal inks applied to temperature-sensitive substrates. The high sintering temperature and easy oxidation limited the application of Cu-based pastes. A two-step method involving liquid co-reduction and heat ripening was developed to synthesize Cu@Sn-Bi core-shell particles. The thickness of Sn-Bi shells can be flexibly adjusted via changing the mass ratio of Cu to Sn-Bi. The volume resistivity of printed circuits using Cu@Sn-Bi pastes solidified at 200 °C was as low as 481 µΩ cm, which increased by 11.8% after an aging process at 190 °C for 6 h. The outstanding stability in a harsh environment would attribute to the effective protection of Sn-Bi alloy shells. This work suggests a new pathway toward the low-temperature bonding and anti-oxidation of Cu particles as conductive fillers, which can be widely applied to the additive manufacturing of flexible wearable electronics.

Programmable mismatch-fueled high-efficiency DNA signal converter.

Herein, by directly introducing mismatched reactant DNA, high-reactivity and high-threshold enzyme-free target recycling amplification (EFTRA) is explored. The developed high-efficiency EFTRA (HEEFTRA) was applied as a programmable DNA signal converter, possessing higher conversion efficiency than the traditional one with perfect complement owing to the more negative reaction standard free energy (ΔG). Once traces of input target miRNA interact with the mismatched reactant DNA, amounts of ferrocene (Fc)-labeled output DNA could be converted via the EFTRA. Impressively, the Fc-labeled output DNA could be easily captured by the DNA tetrahedron nanoprobes (DTNPs) on the electrode surface to form triplex-forming oligonucleotide (TFO) at pH = 7.0 for sensitive electrochemical signal generation and the DTNPs could be regenerated at pH = 10.0, from which the conversion efficiency (N) will be accurately obtained, benefiting the selection of suitable mismatched bases to obtain high-efficiency EFTRA (HEEFTRA). As a proof of concept, the HEEFTRA as an evolved DNA signal converter is successfully applied for the ultrasensitive detection of miRNA-21, which gives impetus to the design of other signal converters with excellent efficiency for ultimate applications in sensing analysis, clinical diagnosis, and other areas.

A universal converting strategy based on target-induced DNA nanoprobe conformational change for lead (II) ion assay.

In this work, an electrochemical biosensor combining a novel enzyme-free converting strategy and branched hybridization chain reaction (bHCR) signal amplification for sensitive detection of Pb2+ is constructed. Herein, inspired by Holliday junctions nanostructure, a relatively symmetric and stable four-way junction nanostructure probe was prepared by using four DNA stands hybridization including P, S1, S2 and help DNA (hD) with rational design and immobilized onto Au@Fe3O4 for rapid separation. With unique advantage of nanoprobe, target Pb2+ will induce the probe conformational change and release S1, S2 from the Au@Fe3O4 scaffold to solution, realizing the conversion of input one target Pb2+ into two DNA outputs with enzyme-free. To further improve the performance of biosensor, bHCR can be triggered by using S1 and S2 as initiator simultaneously to form reticulated dsDNA nanostructure on the surface of electrode. Then, methylene blue (MB) as electron mediator was embedded into the reticulated dsDNA nanostructure to produce a detection signal. This method introduced a universal converting strategy that molecular unrelated to nucleic acids trigger nucleic acids amplification technology with the help of enzyme-free, which not only widen the application of nucleic acids amplification technology, but also has benefited for molecular analysis.

A dynamic, ultra-sensitive and "turn-on" strategy for fluorescent detection of uranyl based on DNAzyme and entropy-driven amplification initiated circular cleavage amplification.

A uranyl detection strategy with ultra-sensitivity was developed based on entropy-driven amplification and DNAzyme circular cleavage amplification. The cleavage of UO22+-specific DNAzyme produces a DNA fragment to initiate the entropy-driven amplification. Two DNA sequences released from the entropy-driven amplification are partly complementary. They can form an entire enzyme strand (E-DNA) of Mg2+-specific DNAzyme. The formed E-DNA can circularly cleave FAM-labeled probes on gold nanoparticles (AuNPs), causing the leaving of FAM from AuNPs and recovery of fluorescent signal. A linear relationship was obtained in the range from 30 pM to 5 nM between fluorescence intensity and concentration of UO22+. The limit of detection was low to 13 pM. This method showed a promising future for practical application in real water samples.

An enzyme-free electrochemical biosensor combining target recycling with Fe3O4/CeO2@Au nanocatalysts for microRNA-21 detection.

In this study, an electrochemical biosensor was proposed for microRNA-21 detection based on Fe3O4/CeO2 @Au magnetite nanoparticles (Fe3O4/CeO2 @Au MNPs) as nanocatalyst and catalytic hairpin assembly (CHA) for signal application. Firstly, target microRNA-21 hybridized with hairpin H2 to form H2-T duplex stranded DNA (dsDNA), which could further open the hairpin H1 for the formation of H1-H2 dsDNA. Simultaneously, the Fe3O4/CeO2 @Au-S1 not only hybridized with single stranded fragment of H1-H2 dsDNA with producing long dsDNA to absorb a large amount of electroactive substances of methylene blue (MB), but also acted as nanocatalyst to directly catalyze the reduction of MB for amplifying the electrochemical signal. Herein, compared with pure Fe3O4 nanoparticles, Fe3O4/CeO2 @Au MNPs exhibited excellent catalytic performance since the cerium oxide (CeO2) nanoparticles and Au nanoparticles can greatly improve the catalytic activity of Fe3O4 nanoparticles and effectively prevent the agglomeration of Fe3O4 nanoparticles. Owing to the signal amplification strategy, the proposed biosensor provided a wide linear range of 1 fM to 1 nM with a low detection limit of 0.33 fM (defined as S/N = 3) for microRNA-21 detection, and exhibited excellent specificity and sensitivity. This strategy provided a novel avenue for the detection of other biomarkers in electrochemical biosensors.

Highly sensitive electrochemical nuclear factor kappa B aptasensor based on target-induced dual-signal ratiometric and polymerase-assisted protein recycling amplification strategy.

In this work, an amplified electrochemical ratiometric aptasensor for nuclear factor kappa B (NF-κB) assay based on target binding-triggered ratiometric signal readout and polymerase-assisted protein recycling amplification strategy is described. To demonstrate the effect of "signal-off" and "signal-on" change for the dual-signal electrochemical ratiometric readout, the thiol-hairpin DNA (SH-HD) hybridizes with methylene blue (MB)-modified protection DNA (MB-PD) to form capture probes, which is rationally introduced for the construction of the assay platform. On the interface, the probes can specifically bind to target NF-κB and expose a toehold region which subsequently hybridizes with the ferrocene (Fc)-modified DNA strand to take the Fc group to the electrode surface, accompanied by displacing MB-PD to release the MB group from the electrode surface, leading to the both "signal-on" of Fc (IFc) and "signal-off" of MB (IMB). In order to improve the sensitivity of the electrochemical aptasensor, phi29-assisted target protein recycling amplification strategy was designed to achieve an amplified ratiometric signal. With the above advantages, the prepared aptasensor exhibits a wide linear range of 0.1pgmL-1 to 15ngmL-1 with a low detection limit of 0.03pgmL-1. This strategy provides a simple and ingenious approach to construct ratiometric electrochemical aptasensor and shows promising potential applications in multiple disease marker detection by changing the recognition probe.

Highly sensitive electrochemical assay for Nosema bombycis gene DNA PTP1 via conformational switch of DNA nanostructures regulated by H+ from LAMP.

The portable and rapid detection of biomolecules via pH meters to monitor the concentration of hydrogen ions (H+) from biological reactions (e.g. loop-mediated isothermal amplification, LAMP) has attracted research interest. However, this assay strategy suffered from inherent drawback of low sensitivity, resulting in great limitations in practical applications. Herein, a novel electrochemical biosensor was constructed for highly sensitive detection of Nosema bombycis gene DNA (PTP1) through transducing chemical stimuli H+ from PTP1-based LAMP into electrochemical output signal of electroactive ferrocene (Fc). With use of target PTP1 as the template, the H+ from LAMP induced the conformational switch of pH-responsive DNA nanostructures (DNA NSs, Fc-Sp@Ts) that was assembled by the hybridization of Fc-labeled signal probe (Fc-Sp) with DNA-based receptor (Ts). Due to the folding of Ts into stable triplex structure at decreased pH, the configuration change of Fc-Sp@Ts led to the releasing of Fc-Sp, which was subsequently immobilized in the electrode interface through the hybridization with the capture probe modified with -SH (SH-Cp), generating amplified electrochemical signal from Fc. The developed biosensor for PTP1 exhibited a reliable linear range of 1 fg µL-1 to 50 ng µL-1 with the limit of detection of 0.31 fg µL-1. Thus, by the regulation of H+ from LAMP reaction on DNA NSs allostery, this novel and simple transduction scheme would be interesting and promising to open up a novel analytical route for sensitive monitoring of different target DNAs in related disease diagnosis.

Amplified impedimetric aptasensor combining target-induced DNA hydrogel formation with pH-stimulated signal amplification for the heparanase assay.

Herein, a novel electrochemical impedimetric biosensor for the heparanase (HPA) assay was developed based on target protein-induced DNA hydrogel formation, followed by pH-stimulation of the hydrogel density to increase the signal amplification. The method involved the synthesis of two different copolymer chains, consisting of two cooperatively functioning cross-linking elements, where one element was associated with the HPA-response and the other one with the pH-response. Initially, single-strand DNA as a capture probe was modified on the electrode surface. In the presence of HPA, the HPA-responsive element binding to HPA-induced DNA hybridization between the two copolymer chains and captured DNA, giving rise to the formation of a low-density polymer hydrogel film on the electrode surface and it obtaining an obvious impedimetric response. A significant signal enhancement was observed when changing the pH of the hydrogel film to 5.0, which could be ascribed to the fact that the pH-responsive element can fold into four-stranded i-motif structures at pH 5.0, leading to the increase in density of the hydrogel film. By implementing the DNA hydrogel to induce an impedimetric response change, this impedimetric biosensor exhibited an excellent analytical performance towards the HPA quantitative assay, with a low detection limit of 0.003 pg mL-1. This new method provides a versatile signal amplification method and paves a new way to construct impedimetric sensors for bioassays.

Thrombin aptasensor enabled by Pt nanoparticles-functionalized Co-based metal organic frameworks assisted electrochemical signal amplification.

In this work, a Pt nanoparticles-functionalized Co-based metal organic frameworks (PtNPs@Co(II)MOFs@PtNPs) was synthesized and applied in electrochemical aptasensor for thrombin (TB) detection. First, the Co(II)MOFs@PtNPs were prepared via the mixed solvothermal method, which consists of inner Pt nanoparticles (PtNPs) encapsulated by aminofunctionalized Co(II)MOFs materials. Following that, additional PtNPs were adsorbed on the surface of Co(II)MOFs@PtNPs, resulting in the formation of PtNPs@Co(II)MOFs@PtNPs nanocomposite. The PtNPs@Co(II)MOFs@PtNPs nanocomposites with a large surface area were implimented as nanocarriers to immobilize a mass of TBA II for the formation of the TBA II bioconjugates that could be captured onto the electrode surface by sandwich-type format. Moreover, the PtNPs@Co(II)MOFs@PtNPs nanocomposites could directly use as redox tags for charge-generating and electron-transporting with the electron transfer from Co(II) to Co(III). Furthermore, in the presence of H2O2, the PtNPs@Co(II)MOF@PtNPs could effectively catalyze H2O2 oxidation with improvement electron transfer of redox probe, resulting in electrochemical signal amplification. Based on the above superior advantages, TB was determined in the concentration range from 0.1pM to 50nM with a detection limit of 0.33fM. Furthermore, the excellent sensitivity and selectivity can be easily established for quantitative analysis of other analytes.

Work-family conflict and safety participation of high-speed railway drivers: Job satisfaction as a mediator.

Despite the large body of work on the work-family interface, hardly any literature has addressed the work-family interface in safety-critical settings. This study draws from social exchange theory to examine the effect of employees' strain-based work-to-family conflict on their supervisors' rating of their safety participation through job satisfaction. The sample consisted of 494 drivers from a major railway company in China. The results of a structural equation model revealed that drivers' strain-based work-to-family conflict negatively influences safety participation, and the relationship was partially mediated by job satisfaction. These findings highlight the importance of reducing employees' work-to-family conflict in safety-critical organizations.

A nanohybrid of platinum nanoparticles-porous ZnO-hemin with electrocatalytic activity to construct an amplified immunosensor for detection of influenza.

In this work, a nanohybrid of platinum nanoparticles-porous ZnO spheres-hemin (Pt-pZnO-hemin) was synthesized for construction of alkaline phosphatase-based immunosensor for detection of influenza. Briefly, porous ZnO spheres (pZnO) were prepared using soluble starches as the capping agent, followed by surface functionalization of platinum nanoparticles via a hydrothermal method (Pt-pZnO). Then, hemin with carboxylic functionality was spontaneously adsorbed onto Pt-pZnO by ester-like binding between carboxylic group of hemin and ZnO. Compared with platinum nanoparticles and hemin, the resulting Pt-pZnO-hemin nanohybrid showed more excellent electrocatalysis activity toward 1-naphthol (1-NP). Taking advantage of the Pt-pZnO-hemin, we have developed an amplified electrochemical immunosensor based on in situ generation of redox probe by alkaline phosphatase (ALP) and Pt-pZnO-hemin as signal enhancer. Herein, electrochemically active 1-NP was generated by enzymatic hydrolysis of inactive 1-naphthyl phosphate by ALP, then Pt-pZnO-hemin was used as catalyst to catalytically oxidize 1-NP, resulting in electrochemical signal amplification. Furthermore, in comparison with other nanomaterials including Au-pZnO, Pt-pZnO and Au-pZnO-hemin, the excellent catalytical property of Pt-pZnO-hemin make it a promising nanohybrid material for ALP-based immunosensor for signal amplification.

Ultrasensitive electrochemical immunosensor for carbohydrate antigen 19-9 using Au/porous graphene nanocomposites as platform and Au@Pd core/shell bimetallic functionalized graphene nanocomposites as signal enhancers.

A facile and feasible sandwich-type electrochemical immunosensor for ultrasensitive determination of Carbohydrate antigen 19-9 (CA19-9) was designed by using Au nanoparticles functionalized porous graphene (Au-PGO) as sensing platform and Au@Pd core/shell bimetallic functionalized graphene nanocomposites (Au@Pd-Gra) as signal enhancers. Herein, Au@Pd-Gra with a large surface area was prepared for immobilizing plentiful of redox probe-thionine (Thi), horseradish peroxidase (HRP) and secondary antibodies (Ab2), leading to the formation of Au@Pd-Gra/Thi-Ab2/HRP bioconjugate which exhibited satisfying electrochemical redox activity, high electrocatalytic activity and friendly biocompatibility. With the synergistic effect between Au@Pd-Gra and HRP, almost triple amplified detection signal was achieved in the presence of H2O2, so as to improve the detection limit of the proposed immunosensor effectively. Furthermore, Au-PGO was utilized as the biosensor platform which could greatly enhance the surface area to immobilize a large amount of captured primary antibodies (Ab1) leading a further enhancement in the sensitivity of immunosensor. Under optimal conditions, the electrochemical immunosensor exhibited desirable performance for determination of CA19-9 with a wide linearity in the range from 0.015 to 150UmL(-1) and a relatively low detection limit of 0.006UmL(-1). Importantly, the resulted immunosensor displayed good specificity and high sensitivity, implying potential applications in clinical research.

Amplified thrombin aptasensor based on alkaline phosphatase and hemin/G-quadruplex-catalyzed oxidation of 1-naphthol.

An alkaline phosphatase (ALP)-based biosensor can in situ generate an electroactive product by enzymatic hydrolysis of inactive substrates. To obtain a higher signal-to-background ratio, a chemical redox cycling signal-amplified strategy based on the addition of a strong reducing agent has often be applied in the construction of ALP-based biosensors. However, the strong reducing agent not only affects the activity of ALP but also readily reacts with dissolved oxygen, leading to inaccurate results. In this work, a new signal-amplified strategy for a thrombin (TB) aptasensor based on the catalytic oxidation of ALP-generated products, 1-naphthol (NP), using hemin/G-quadruplex DNAzymes was reported. We implemented gold-nanoparticle-decorated zinc oxide nanoflowers (Au-ZnO) as the matrix for immobilizing ALP and TB aptamer (TBA) and then labeled it with hemin to form hemin/G-quadruplex/ALP/Au-ZnO bioconjugates (TBA II bioconjugates). Through a "sandwich" reaction, TBA II bioconjugates were captured on the electrode surface. The amplified signal was carried out in two steps: (i) an ALP-catalyzed inactive substrate, 1-naphthyl phosphate (NPP), in situ produces NP on the surface of the electrode; (ii) on the one hand, NP as a new reactant could be directly electrooxidized and generated an electrochemical signal, but, on the other hand, NP could be oxidized by hemin/G-quadruplex in the presence of H2O2, resulting in amplification of the electrochemical signal. The proposed TB aptasensor achieved a linear range of 1 pM to 30 nM with a detection limit of 0.37 pM (defined as S/N = 3).