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Abstract Along with the advance in medical research, more biomarkers emerge as useful indicators for both disease and health index. However, majority of them have no practical or economic testing methods available yet, or rely on high-costing methods such as Enzyme-Linked Immuno-Sorbent Assay (ELISA), High-Performance Liquid Chromatography (HPLC), Mass Spectrum, and immunohistochemistry (IHC). In this article, we develop a universal nanozyme integrated testing platform for biological molecules that incorporates the electrochemical measurement of glucose with lateral flow immunostrip (LFS) for target analytes. This design involves the quantitative conversion of analytes into invertase and then glucose, which can be measured by an extremely affordable meter. The feasibility of this design was validated using 8-hydroxy-2'-deoxyguanosine (8-OHdG) and prostate specific antigen (PSA) as representatives for small molecules and moderate to large proteins respectively. Our approach yields results comparable to commercial diagnostic ELISA kits at a substantially reduced cost and reaction time. Specifically, the design has a detection limit of 0.23 ng mL-1 for 8-OHdG and 1.26 ng mL-1 for PSA, and a detection range of 0.1–100 ng mL-1 for 8-OHdG and 1–100 ng mL-1 for PSA. By combining the accessibility of well-established glucose testing and LFS, our design can serve as a point of care testing method that can be fully integrated into the personal lifestyle without requiring professional assistance. Highlights • Developed a universal nanozyme integrated lateral fluidic sensing platform. • Non-glucose biomarkers can be measured by a glucose meter based on this platform. • Two types of molecules were successfully detected through this platform. • The sensor was easy to fabricate and convenient to operate. • This approach will provide new opportunities for the detection of other targets. [ABSTRACT FROM AUTHOR]

The accurate therapeutic evaluation for chronic myeloid leukemia (CML) drug is of great importance to minimize side effects and enhance efficacy. Herein, a facile and precise surface-enhanced scattering (SERS) approach based on coupled plasmonic field has been introduced to evaluate the therapeutic outcomes of antileukemia drug through ultrasensitive assay of caspase-3 activity in apoptotic cells. Caspase-3 as an apoptosis indicator could specifically cleave the N-terminus of biotinylated DEVD-peptide (biotin-Gly-Asp-Gly-Asp-Glu-Val-Asp-Gly-Cys) immobilized on the Au nanoparticle-decorated TiO 2 nanotube arrays (TiO 2 /Au NTAs) substrate. After the enzyme cleavage with caspase-3, Raman-labelled Au nanostar (AuNS) probes captured the residual DEVD-peptides via the recognition between streptavidin and biotin, thus resulting in an enhanced Raman response on the SERS platform. The variation of Raman intensity revealed caspase-3 activity that reflected the chemotherapeutic effect. On this platform, AuNS nanoprobes offered a large number of binding sites and intrinsic “hot spots” for Raman reporters, while TiO 2 /Au NTAs rendered a homogenously coupled electromagnetic field between the adjacent repeated units over the large area. In particular, a spatially expanding plasmonic field formed by coupling AuNSs with TiO 2 /Au NTAs would further heighten Raman enhancement. Taking these advantages, the strong and uniform Raman signals were achieved. Furthermore, the practicability investigation witnessed that the proposed SERS strategy was available to evaluate the therapeutic effect of dasatinib on CML K562 cells. The developed method possesses fascinating advantages of cost-effectiveness, excellent reproducibility and high sensitivity, which endows it with promising potential in apoptosis monitoring and anticancer drug development. [ABSTRACT FROM AUTHOR]

Sensitive and specific detection of DNA is of great significance for clinical diagnosis. In this paper, an effective cascade signal amplification strategy was introduced into photoelectrochemical (PEC) biosensor for ultrasensitive detection of human T-cell lymphotropic virus type I (HTLV-I) DNA. This proposed signal amplification strategy integrates λ-exonuclease (λ-Exo) aided target recycling with hybridization chain reaction (HCR) and enzyme catalysis. In the presence of target DNA (tDNA) of HTLV-I, the designed hairpin DNA (h 1 DNA) hybridized with tDNA, subsequently recognized and cleaved by λ-Exo to set free tDNA. With the λ-Exo aided tDNA recycling, an increasing number of DNA fragments (output DNA, oDNA) were released from the digestion of h 1 DNA. Then, triggered by the hybridization of oDNA with capture DNA (cDNA), numerous biotin-labeled hairpin DNAs (h 2 DNA and h 3 DNA) could be loaded onto the photoelectrode via the HCR. Finally, avidin-labeled alkaline phosphatase (avidin-ALP) could be introduced onto the electrode by specific interaction between biotin and avidin. The ALP could catalyze dephosphorylation of phospho-L-ascorbic acid trisodium salt (AAP) to generate an efficient electron donor of ascorbic acid (AA), and thereby greatly increasing the photocurrent signal. By utilizing the proposed cascade signal amplification strategy, the fabricated PEC biosensor exhibited an ultrasensitive and specific detection of HTLV-I DNA down to 11.3 aM, and it also offered an effective strategy to detect other DNAs at ultralow levels. [ABSTRACT FROM AUTHOR]

In the present work, electrogenerated chemiluminescence (ECL) of luminol was investigated in neutral condition at a gold electrode in the presence of silicon quantum dots (SiQDs). The results revealed that SiQDs can not only greatly enhance luminol ECL, but also act as energy acceptor to construct a novel ECL resonance energy transfer (ECL-RET) system with luminol. As a result, strong anodic ECL signal was obtained in neutral condition at the bare gold electrode, which is suitable for biosensing application. Lysozyme exhibited apparent inhibiting effect on the ECL-RET system, based on which an ECL aptasensor was fabricated for the sensitive detection of lysozyme. The proposed method showed high sensitivity, good selectivity, and wide linearity for the detection of lysozyme in the range of 5.0×10 –14 −5.0×10 −9 g mL −1 with a detection limit of 5.8×10 –15 g mL −1 (3σ). The results suggested that as-proposed luminol/SiQDs ECL biosensor will be promising in the detection enzyme. [ABSTRACT FROM AUTHOR]

An ultrasensitive electrochemiluminescence (ECL) biosensor for the detection of microRNA was developed based on nicking enzymes Nb.BbvCI mediated signal amplification (NESA). First, the hairpin probe1-N-CQDs with assistant probe and microRNA (miRNA) formed Y junction structure which was cleaved with the addition of nicking enzymes Nb.BbvCI to release miRNA and assistant probe. Subsequently, the released miRNA and assistant probe can initiate the next recycling process. The generation of numerous intermediate sequences nitrogen doped carbon quantum dots-DNA (N-CQDs-DNA) can further hybridize with hairpin probe2 immobilized on GO/Au composite modified electrode surface, the initial ECL intensity was enhanced. The ECL intensity would increase with increasing concentration of the target miRNA, and the sensitivity of biosensor would be promoted because of the efficient signal amplification of the target induced cycling reaction. The novel designed biosensor provided a highly sensitive and selective detection of miRNA-21 from 10 aM to10 4 fM with a relatively low detection limit of 10 aM. Thus, our strategy has a potential application in the clinical diagnosis. [ABSTRACT FROM AUTHOR]

Electrogenerated chemiluminescence (ECL) of semiconductor quantum dots (QDs) has been considered as a powerful technique in the fabrication of biosensor, however, high-toxicity of heavy metal ion containing in QDs severely limits their further applications, and the search for the alternative benign nanomaterials with high ECL efficiency is urgent. Herein, ECL behavior of eco-friendly silicon quantum dots (SiQDs) was reported in neutral aqueous condition. Stable and intense cathodic ECL emission was obtained in phosphate buffer solution (PBS) with K 2 S 2 O 8 as coreactant. ECL resonance energy transfer (ECL-RET) system was established with SiQDs ECL as energy donor and gold nanoparticles (AuNPs) as energy acceptor, based on which a novel ECL biosensor was fabricated. AuNPs was connected at the terminal of hairpin DNA to form a signal probe. When the probe was modified on SiQDs, ECL-RET occurred due to the short distance between AuNPs and SiQDs, resulting in the apparent decrease of ECL signal. Target DNA can open the loop of hairpin DNA, and move AuNPs away from the electrode surface. As a result, the ECL-RET process was hampered, and the ECL emission resumed. The increased ECL signals varied linearly with the target DNA concentrations in the range of 0.1 fM to 1 pM with the detection limit of 0.016 fM (3σ). The proposed ECL sensor exhibited highly sensitivity and good selectivity in the detection of target DNA. [ABSTRACT FROM AUTHOR]

Quantifying the glycan expression status on cell surfaces is of vital importance for insight into the glycan function in biological processes and related diseases. Here we developed a versatile aptasensor for electrochemical quantification of cell surface glycan by taking advantage of the cell-specific aptamer, and the lectin-functionalized gold nanoparticles acting as both a glycan recognition unit and a signal amplification probe. To construct the aptasensor, amine-functionalized mucin 1 protein (MUC1) aptamer was first covalently conjugated to carboxylated-magnetic beads (MBs) using the succinimide coupling (EDC-NHS) method. On the basis of the specific recognition between aptamer and MUC1 protein that overexpressed on the surface of MCF-7 cells, the aptamer conjugated MBs showed a predominant capability for cell capture with high selectivity. Moreover, a lectin-based nanoprobe was designed by noncovalent assembly of concanavalin A (ConA) on gold nanoparticles (AuNPs). This nanoprobe incorporated the abilities of both the specific carbohydrate recognition and the signal amplification based on the gold-promoted reduction of silver ions. By coupling with electrochemical stripping analysis, the proposed sandwich-type cytosensor showed an excellent analytical performance for the ultrasensitive detection of MCF-7 cells and quantification of cell surface glycan. More importantly, taking advantage of Con A-gold nanoprobe catalyzed silver enhancement, the proposed method was further used for naked-eye tracking glycolytic inhibition in living cells. This aptasensor holds great promise as a new point-of-care diagnostic tool for analyzing glycan expression on living cells and further helps cancer diagnosis and treatment. [ABSTRACT FROM AUTHOR]

MicroRNA-21 (miR-21) is a promising diagnostic biomarker for breast cancer screening and disease progression, thus the method for the sensitive and selective detection of miR-21 is vital to its clinical diagnosis. Herein, we develop a novel method to quantify miR-21 levels as low as attomolar sensitivity by a target-triggered triple isothermal cascade amplification (3TICA) strategy. An ingenious unimolecular DNA template with three functional parts has been designed: 5′-fragment as the miR-21 recognition unit, middle fragment as the miR-21 analogue amplification unit, and 3′-fragment as the 8–17 DNAzyme production unit. Triggered by miR-21 and accompanied by polymerase-nicking enzyme cascade, new miR-21 analogues autonomously generated for the successive re-triggering and cleavage process. Simultaneously, the 8–17 DNAzyme-contained sequence could be exponentially released and activated for the second cyclic cleavage toward a specific ribonucleotide (rA)-contained substrate, inducing a remarkably amplified generation of HRP-mimicking DNAzyme in the presence of hemin. Finally, the amperometric technique was used to record the catalytic reduction current of 3,3′,5,5′-tetramethylbenzidine (TMB) in the presence of H 2 O 2 . The increase in the steady-state current was proportional with the increase of the miR-21 concentration from 1 aM to 100 pM. An ultra-low detection limit of 0.5 aM with an excellent selectivity for even discriminating differences between 1-base mismatched target and miR-21 was achieved. This simple and cost-effective 3TICA strategy is promising for the detection of any short oligonucleotides, simply by altering the target recognition unit in the template sequence. [ABSTRACT FROM AUTHOR]

Since atherosclerosis, a disease characterized by abnormal arterial lipid deposition, may lead to fatal cardiovascular diseases, imaging of atherosclerotic plaques is of great value for their pathological assessment. In this study, we propose a lipid droplet (LD)-hitchhiking strategy to in situ create Trojan foam cells for fluorescence/photoacoustic imaging of atherosclerotic plaques via homologous targeting effect. In our design, functional liposomes (DCP liposomes) composed of phospholipid dioleoylphosphatidylserine (DOPS), a novel LD inducer we found, and Cypate-PC, a synthesized lipid-like molecular probe, have demonstrated great capability of inducing LDs in monocytes/macrophages while being enveloped into the resulting Trojan foam cells. Taking advantage of homologous targeting effect, the imaging probe hitchhikes on the LDs in Trojan foam cells for targeted transport to the plaque sites. Moreover, the confinement in highly hydrophobic LDs endows the imaging probe with high efficiency in light absorption, enabling greatly intensified fluorescence/photoacoustic signals. The DCP liposomes have shown great potency in inducing the generation of Trojan foam cells, and eventually ex vivo fluorescence imaging and in vivo photoacoustic imaging of atherosclerotic plaques. The proposed strategy provides more insights into the design of targeted imaging methodologies, and also an effective avenue to facilitate the evaluation and subsequent treatment of atherosclerotic plaques. • In situ lipid droplets formation in monocytes/macrophages (endocytosis of lipid). • Homologous targeting of Trojan foam cells to foam cells in atherosclerotic plaques. • Imaging probes hitchhiking on LDs in Trojan foam cells for atherosclerotic plaque targeting. • Hydrophobic LDs' environment elevating the light absorption efficiency of probes for combined FL/PA imaging. [ABSTRACT FROM AUTHOR]

A novel, enhanced photoelectrochemical immunoassay was established for sensitive and specific detection of carbohydrate antigen 19-9 (CA19-9, Ag). In this protocol, TiO 2 nanowires (TiO 2 NWs) were first decorated with Au nanoparticles to form TiO 2 NWs/Au hybrid structure, and then coated with CdSe@ZnS quantum dots (QDs) via the layer-by-layer method, producing TiO 2 NWs/Au/CdSe@ZnS sensitized structure, which was employed as the photoelectrochemical matrix to immobilize capture CA19-9 antibodies (Ab 1 ); whereas, bipyridinium (V 2+ ) molecules were labeled on signal CA19-9 antibodies (Ab 2 ) to form Ab 2 @V 2+ conjugates, which were used as signal amplification elements. The TiO 2 NWs/Au/CdSe@ZnS sensitized structure could adequately absorb light energy and dramatically depress electron–hole recombination, resulting in evidently enhanced photocurrent intensity of the immunosensing electrode. While target Ag were detected, the Ab 2 @V 2+ conjugates could significantly decrease the photocurrent detection signal because of strong electron-withdrawing property of V 2+ coupled with evident steric hindrance of Ab 2 . Thanks to synergy effect of TiO 2 NWs/Au/CdSe@ZnS sensitized structure and quenching effect of Ab 2 @V 2+ conjugates, the well-established photoelectrochemical immunoassay exhibited a low detection limit of 0.0039 U/mL with a wide linear range from 0.01 U/mL to 200 U/mL for target Ag detection. This proposed photoelectrochemical protocol also showed good reproducibility, specificity and stability, and might be applied to detect other important biomarkers. [ABSTRACT FROM AUTHOR]

A novel “ON–OFF” switchable enzymatic biofuel cell (EBFC), controlled by in situ thermal-stimuli signal, has been consciously designed. Poly ( N -isopropylacrylamide) (PNIPAm) chains were used to act as “ON” and “OFF” channels. Consecutively switching of temperature below and above the lower critical solution temperature (LCST), the reversible conformation changing of the PNIPAm chains between superhydrophilicity and superhydrophobicity was achieved, which constructed the “ON” and “OFF” channel for the transfer of the electrochemical probe to the underlying electrode correspondingly. Gold nanoparticles (AuNPs) protected glucose oxidase and laccase were successfully entrapped into the intelligent thermal-sensitive PNIPAm chains, and performed as the catalysts for the oxidation of glucose and the reduction of O 2 , respectively. Below the LCST, the fuels and the mediator could access to the catalytic centers of enzymes (set as “ON” state); while above the LCST, the reaction was impeded because the process of reactant transmission was blocked (set as “OFF” state). By switching the “valve” of mass transfer, the fabricated EBFC displayed the obvious “ON–OFF” controllable behavior. At the “ON” state, the open circuit voltage ( E cell ocv ) and maximal power output density ( P max ) could reach to 0.70 V and 20.52 μW cm −2 , respectively; while at the “OFF” state, the E cell ocv and P max were only 0.30 V and 3.28 μW cm −2 correspondingly. The switchable process was repeatable, and the response time was only several minutes. [ABSTRACT FROM AUTHOR]

A triple-signal amplification strategy was proposed for highly sensitive and selective detection of thymine DNA glycosylase (TDG) by coupling a dendrimer-like DNA label with the electrochemical method and quantum dots (QDs) tagging. The DNA-QDs dendrimer-like superstructure was designed by DNA hybridization and covalent assembling. Benefiting from outstanding performance of the amplification strategy, this assay showed high sensitivity, extraordinary stability, and easy operation. The limit of detection could reach 0.00003 U µL −1 with a splendid specificity. The TDG content in different concentration of HeLa cell was also determined. This assay opens a new horizon for both qualitative and quantitative detection of TDG, holding great promise for potential application in cancer cell research and clinical diagnostics. [ABSTRACT FROM AUTHOR]

A novel electrochemical biosensing platform for nicotinamide adenine dinucleotide (NAD+)-dependent dehydrogenase catalysis was designed using the nitrogen-doped graphene (NG), which had properties similar to NADH dehydrogenase (CoI). NG mimicked flavin mononucleotide (FMN) in CoI and efficiently catalyzed NADH oxidation. NG also acted as an electron transport "bridge" from NADH to the electrode due to its excellent conductivity. In comparison with a bare gold electrode, an 800mV decrease in the overpotential for NADH oxidation and CoI-like behavior were observed at NG-modified electrode, which is the largest decrease in overpotential for NADH oxidation reported to date. The catalytic rate constant (k) for the CoI-like behavior of NG was estimated to be 2.3×105M−1s−1, which is much higher than that of other previously reported FMN analogs. The Michaelis-Menten constant (Km) of NG was 26μM, which is comparable to the Km of CoI (10μM). Electrodes modified with NG and NG/gold nanoparticals/formate dehydrogenase (NG/AuNPs/FDH) showed excellent analytical performance for the detection of NADH and formate. This electrode fabrication strategy could be used to create a universal biosensing platform for developing NAD+-dependent dehydrogenase biosensors and biofuel cells. [ABSTRACT FROM AUTHOR]

In recent decades, advanced therapies and novel scientific drug evaluation systems for chronic myeloid leukemia (CML) treatment are very urgent due to its increasing morbidity. The combination of dasatinib with tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) was supposed to be effective for leukemia therapy. Taking full advantage of novel nano-biotechnology, we have developed a robust electrochemical cytosensing approach to profile the therapeutic effects of dasatinib and TRAIL by probing the activity of caspase-3 from apoptotic CML cells. The sensor was on a base of a glassy carbon electrode (GCE) modified with nano-materials composed of Au nanoparticles (AuNPs), poly(dimethyl diallyl ammonium chloride) (PDDA), and carbon nanotubes (CNTs). Then the platform immobilized the biotinylated DEVD-peptide (biotin-Gly-Asp-Gly-Asp-Glu-Val-Asp-Gly-Cys) via the strong bonding between AuNPs and the thiol group (Au-S bond). In particular, the sensor was then constructed with the environmentally friendly alkaline phosphatase (ALP) via the specific interaction between the biotin and streptavidin, and could retest detection indirectly for caspase-3 sensing by detecting the differential pulse voltammetry (DPV) signal of enzymatic catalysis product, ascorbic acid (AA). The results indicated that either dasatinib or TRAIL could successfully induce the apoptosis of CML cells, while the combination of dasatinib and TRAIL resulted in an improved therapeutic effect, suggesting a novel optimized strategy for CML therapy. This novel electrochemical sensing strategy exhibits attractive advantages of environmental benignity, simple performance, high stability, and may be readily expanded to evaluate other cancer therapeutic effects. [ABSTRACT FROM AUTHOR]

Abstract: Dual co-sensitized structure of TiO2/CdS/CdSe was designed to develop a novel photoelectrochemical immunoassay for highly sensitive detection of human interleukin-6 (IL-6). To construct a sensing electrode, TiO2/CdS hybrid was prepared by successive adsorption and reaction of Cd2+ and S2− ions on the surface of TiO2 and then was employed as matrix for immobilization of anti-IL-6 antibody, whereas CdSe QDs linked to IL-6 were used for signal amplification via the specific antibody–antigen immunoreaction between anti-IL-6 and IL-6–CdSe bioconjugate. Greatly enhanced sensitivity for IL-6 detection was derived from the new co-sensitization signal amplification strategy. First, the TiO2/CdS/CdSe co-sensitized structure extended the absorption range to long wavelength of white light, which adequately utilized the light energy. Second, the TiO2/CdS/CdSe co-sensitized structure possessed stepwise band-edge levels favoring ultrafast transfer of photogenerated electrons and significantly prompted the photoelectrochemical performance. Besides, the introduction of CdSe effectively prevented the recombination of photogenerated electrons in the conduction band of CdS, further causing an enhanced photocurrent. Accordingly, upon the co-sensitization strategy, a novel immunoassay based on the competitive binding of anti-IL-6 antibody with IL-6 antigen and IL-6–CdSe bioconjugate was developed, and it exhibited a wide linear range from 1.0pg/mL to 100ng/mL with a low detection limit of 0.38pg/mL for IL-6 detection. The proposed co-sensitization strategy presented high sensitivity, reproducibility, specificity and stability, and also opened up a new promising platform for detection of other biomarkers. [Copyright &y& Elsevier]