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  • Nanoconjugates of ferrocene and carbon-encapsulated iron nanoparticles as sensing platforms for voltammetric determination of ceruloplasmin in blood
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-23
    Jakub P. Sęk, Artur Kasprzak, Michał Bystrzejewski, Magdalena Poplawska, Waldemar Kaszuwara, Zbigniew Stojek, Anna M. Nowicka

    The nanoparticles comprising of iron core and carbon shell were decorated with ferrocene derivatives: ferrocenecarboxaldehyde (Fc-1) and ferrocenecarboxaldehyde oxime (Fc-2). A microdrop of suspension of the nanoconjugate was placed on a glassy-carbon electrode to prepare the recognition/sensing layer. Drying and purification of the sensing layer resulted in a well-defined and stable square-wave voltammogram of the ferrocene moiety. The height of the voltammetric peak increased in the presence of ceruloplasmin. That increase was linearly dependent on the logarithmic concentration of ceruloplasmin in blood. The applied external magnetic field was a factor which yielded better sensitivity and repeatability of the sensor response. The linearity of sensor response was found to be between 0.001 and 10 μg dL−1 and 0.05 to 10 μg dL−1 for both nanoconjugates: Fe@C-Fc-1 and Fe@C-Fc-2, in the presence and absence of the magnet, respectively. The obtained detection limit (LOD) for Fe@C-Fc-1 was found to be 0.60 and 0.10·μg·dL−1 in the absence and presence of magnetic field, respectively, whilst for Fe@C-Fc-2 was 0.4 and 0.07·μg·dL−1 in the absence and presence of a magnet, respectively. The proposed method is selective because the presence of common antioxidants in blood did not interfere significantly with the determination of the concentration of ceruloplasmin.

    更新日期:2017-11-24
  • Biofunctionalized silicon nitride platform for sensing applications
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-23
    Hiofan Hoi, Salva S. Rezaie, Lu Gong, Payel Sen, Hongbo Zeng, Carlo Montemagno, Manisha Gupta

    Silicon nitride (SiNx) based biosensors have the potential to converge on the technological achievements of semiconductor microfabrication and biotechnology. Development of biofunctionalized SiNx surface and its integration with other devices will allow us to integrate the biosensing capability with probe control, data acquisition and data processing. Here we use the hydrogen plasma generated by inductively coupled plasma-reactive ion etching (ICP-RIE) technique to produce amino-functionality on the surface of SiNx which can then be readily used for biomolecule immobilization. ICP-RIE produces high-density hydrogen ions/radicals at low energy, which produces high-density amino group on the SiNx surface within a short duration of time and with minimal surface damage. In this work, we have demonstrated selective amination of SiNx surface as compared to Si surface. The as-activated SiNx surface can be readily biofunctionalized with both protein and oligonucleotide through covalent immobilization. N-5-azido-2-nitrobenzoyloxysuccinimide, a photoactivable amino reactive bifunctional crosslinker, was used and greater than 90% surface coverage was achieved for protein immobilization. In addition, ssDNA immobilization and hybridization with its complemented strand was shown. Thus, we demonstrate a uniform, reliable, fast and economical technique for creating biofunctionalized SiNx surface that can be used for developing compact high-sensitivity biosensors.

    更新日期:2017-11-24
  • Nitration of Tyrosine and Its Effect on DNA Hybridization
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-23
    Seda Nur Topkaya, Vasfiye Hazal Ozyurt, Arif E. Cetin, Semih Otles

    One major marker of nitrosative stress is the formation of 3-Nitrotyrosine (3-NT) from Tyrosine (Tyr) by adding a nitro group (-NO2) with nitrating agents. Nitration of Tyr often causes loss of protein activity and is linked with many diseases. In this article, we detect 3-NT and discriminate it from Tyr with Differential Pulse Voltammetry (DPV) as it is a very important biomarker. We first examined redox (oxidation/reduction) properties and stability of 3-NT in detail. Second, we provided the Tyr and 3-NT discrimination with DPV and compared with the chromatography. We then explored the interaction of 3-NT and DNA oligonucleotides. Our findings demonstrate that 3-NT can be used as a new electrochemical indicator, which is able to detect hybridization of probe (single stranded DNA-ssDNA) and hybrid (double stranded DNA-dsDNA) both via 3-NT reduction and guanine oxidation signal changes at the same time. The signal differences enabled us to distinguish ssDNA and dsDNA without using a label or a tag. Moreover, we achieved to detect hybridization of DNA by using the reduction signal of 3-NT obtained at −0.4 V vs. Ag/AgCl. More importantly, we observed the changes of the reduction signals of 3-NT after the interaction of probe and hybrid sequences. We showed that 3-NT signal decreases more with hybrid than the probe. Our platform, for the first time, demonstrates the detection of hybridization both guanine oxidation and indicator reduction signal changes at the same time. Moreover, we, for the first time, demonstrated the interaction between 3-NT and DNA.

    更新日期:2017-11-23
  • A single-cell analysis platform for electrochemiluminescent detection of platelets adhesion to endothelial cells based on Au@DL-ZnCQDs nanoprobes
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-22
    Dongping Long, Yunfei Shang, Youyi Qiu, Bin Zhou, Peihui Yang

    A novel single-cell analysis platform (SCA) was developed for the investigation of platelets adhesion to single human umbilical vein endothelial cell (HUVEC) via using the adhesion molecule (E-selectin) on the damaged HUVEC as the marker site, and integrating electrochemiluminescence (ECL) with the ultrasensitive Au@DL-ZnCQDs nanoprobes. The Au@DL-ZnCQDs nanocomposite, a kind of double layer zinc-coadsorbed carbon quantum dot (ZnCQDs) core-shell nanoprobe, was firstly constructed by using gold nanoparticles (AuNPs) as the core to load with ZnCQDs and then the citrate-modified silver nanoparticles (AgNPs) as the bridge to link AuNPs-ZnCQDs with ZnCQDs to form the core-shell with double layer ZnCQDs (DL-ZnCQDs) nanoprobe, revealed a 10-fold signal amplification. The H2O2-induced oxidative damage HUVECs were utilized as the cellular model on which anti-E-selectin functionalized nanoprobes specially recognized E-selectin, the SCA showed that the ECL signals decreased with platelets adhesion to single HUVEC. The proposed SCA could effectively and dynamically monitor the adhesion between single HUVEC and platelets in the absence and presence of collagen activation, moreover, be able to quantitatively detect the number of platelets adhesion to single HUVEC, and show a good analytical performance with linear range from 1 to 15 platelets. In contrast, the HUVEC was down-regulated the expression of adhesion molecules by treating with quercetin inhibitor, and the SCA also exhibited the feasibility for analysis of platelets adhesion to single HUVEC. Therefore, the single-cell analysis platform provided a novel and promising protocol for analysis of the single intercellular adhesion, and it will be beneficial to elucidate the pathogenesis of cardiovascular diseases.

    更新日期:2017-11-23
  • Antibody biosensors for spoilage yeast detection based on impedance spectroscopy
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-22
    I. Tubía, J. Paredes, E. Pérez-Lorenzo, S. Arana

    Brettanomyces is a yeast species responsible for wine and cider spoilage, producing volatile phenols that result in off-odors and loss of fruity sensorial qualities. Current commercial detection methods for these spoilage species are liable to frequent false positives, long culture times and fungal contamination. In this work, an interdigitated (IDE) biosensor was created to detect Brettanomyces using immunological reactions and impedance spectroscopy analysis. To promote efficient antibody immobilization on the electrodes’ surface and to decrease non-specific adsorption, a Self-Assembled Monolayer (SAM) was developed. An impedance spectroscopy analysis, over four yeast strains, confirmed our device's increased efficacy. Compared to label-free sensors, antibody biosensors showed a higher relative impedance. The results also suggested that these biosensors could be a promising method to monitor some spoilage yeasts, offering an efficient alternative to the laborious and expensive traditional methods.

    更新日期:2017-11-22
  • PdPt nanoparticles anchored on the N-G with the integration of PANI nanohybrids as novel redox probe and catalyst for the detection of rs1801177
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-21
    Jing Wu, Junlin He, Chengli Zhang, Jun Chen, Yazhen Niu, Qiqi Yuan, Chao Yu

    Single nucleotide polymorphism (SNP) in lipoprotein lipase (LPL) gene (rs1801177) is strongly associated with the increased progression of atherosclerosis, threatening global public health. In this work, a relatively simple, specific and ultrasensitive electrochemical DNA biosensor was constructed to detect rs1801177 for the first time. A glass carbon electrode was modified with fullerene (C60)/polyamidoamine (PAMAM)/gold (Au) nanoparticles nanocomposites film. In addition the nitrogen-doped graphene (N-G)/palladium platinum (PdPt) bimetallic nanoparticle/ polyaniline (PANI) nanohybrids were synthesised and used to label the signal probes. These nanohybrids have abundant active groups, and efficient redox and catalytic activity, allowing them to be used as the nanocarrier for a redox nanoprobe without the additional modification of electroactive substance and catalyst, which could effectively simplify the operation procedure and shorten the analysis time. With the catalysis of H2O2 by nanohybrids, the detection signal of N-G/PdPt/PANI itself could be significantly enhanced, lead to the improvement of the sensitivity. Under optimal conditions, the electrochemical DNA biosensor exhibited desirable performance for the determination of rs1801177 with a wide linearity ranging from 10 fM to 10 nM and a relatively low detection limit of 3.33 fM (S/N=3). The proposed biosensor showed excellent selectivity to the target DNA compared to possible interfering substances. The results suggested that this method has potential applications in clinical research.

    更新日期:2017-11-21
  • Superwettable Microchips with Improved Spot Homogeneity toward Sensitive Biosensing
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-21
    Yanxia Chen, Li-Ping Xu, Jingxin Meng, Shaohui Deng, Lulin Ma, Shudong Zhang, Xueji Zhang, Shutao Wang

    The high-quality spots in microchips are prerequisites for sensitive and accurate detection of biomarkers. In this work, the superwettable micropattern was constructed by introducing superhydrophilic microwells onto a superhydrophobic substrate. The sample can distribute homogeneously within the well-designed superhydrophilic microwells after droplet evaporation and form homogeneous deposit spots, which can be ascribed to the enhanced Marangoni effect in superwettable micropattern and the suppressed outward flow by 3D nanodendritic silica structure. Based on the improved homogeneity of spots, sensitive and accurate fluorescence readout could be obtained. The free prostate-specific antigen (f-PSA) microchip based on the superwettable micropattern was developed. This superwettable f-PSA microchip exhibits high sensitivity, excellent specificity and long-term stability, and a limit of detection as low as 10 fg mL-1 is achieved. Moreover, the superwettable f-PSA microchip can accurately detect human serum samples with excellent correlations with chemiluminescence immunoassay in the clinic, demonstrating its great potential as a sensitive and reliable sensing platform for biological analysis and clinical diagnosis.

    更新日期:2017-11-21
  • Ultrasensitive electrochemiluminescence assay of tumor cells and evaluation of H2O2 on a paper-based closed-bipolar electrode by in-situ hybridization chain reaction amplification
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-21
    Shenguang Ge, Jinge Zhao, Shaopeng Wang, Feifei Lan, Mei Yan, Jinghua Yu

    In this manuscript, a disposable paper-based analytical device comprised of a closed bipolar electrode (BPE) was fabricated for the ultrasensitive electrochemiluminescence (ECL) detection of intracellular H2O2 and the number of cancer cells. In this approach, wax printing was used to fabricated reaction zone, and carbon ink-based BPE and driving electrodes were screen-printed into the paper. AuPd nanoparticles (NPs), which served as a carrier of the capture aptamer and as the catalyst for the ECL reaction of luminol and H2O2, were used to modify the BPE. Luminol/Au NPs were attached to the surface of the captured cells via hybridation chain reaction with two hairpin structure DNA labelled luminol/Au NPs. In the stimulation of phorbol myristate acetate, The coreactant H2O2 was released from the target cells. The ECL response of the luminol-H2O2 system was related to the number of cancer cells in the testing buffer, which served as a quantitative signal for the determination of cancer cells and the concentration of H2O2. In order to decrease the external voltage, K3[Fe(CN)6] was introduced in the cathode resevoir of BPE because it gained electrons at the cathode more easily than oxygen. The ECL intensity was quantitatively related to the concentration of MCF-7 in the range of 1.0×102 to 1.0×107 cells/mL. The detection limit was 40 cells/mL and it showed good specificity for cells with high overexpression of mucin-1 receptor, it was concluded that the developed protocol could be effectively utilized for the detection of MCF-7 cells.

    更新日期:2017-11-21
  • A multi-functional gold/iron-oxide nanoparticle-CNT hybrid nanomaterial as virus DNA sensing platform
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-21
    Jaewook Lee, Masahiro Morita, Kenshin Takemura, Enoch Y. Park

    Nanoparticle (NP)-carbon nanomaterial hybrid structures have garnered significant attention due to their synergistic effects. Based on their interesting properties, hybrid nanomaterial-based sensing platforms have been investigated to achieve high sensitivity and selectivity detection. In our study, binary-NP-decorated carbon nanotubes (bNP-CNTs) were successfully synthesized through a simple two-step method and applied as a biosensing platform. Gold (Au)/iron-oxide magnetic NP-decorated CNTs (Au/MNP-CNT) were used for influenza and norovirus DNA sensing channels. To demonstrate biosensing, first, the Au/MNP-CNTs were magnetically aligned on a Pt-interdigitated electrode, and then, a thiol-group-functionalized probe DNA was attached to the Au NP surface on the bNP-CNT hybrid structure through thiol chemistry. DNA hybridization between the target influenza or norovirus DNA and probe DNA was measured to monitor an electrical conductivity change of the Au/MNP-CNTs. Various concentrations of target DNA from 1 pM to 10 nM were monitored, and the limits of detection for influenza virus and norovirus were calculated to be approximately 8.4 pM and 8.8 pM, respectively. The specificity was confirmed using different mismatched DNA sequences, showing high specificity. Therefore, this Au/MNP-CNT-based DNA sensing system exhibited excellent detection potential, and such hybrid materials could be universally applied as a highly sensitive and selective biosensing system.

    更新日期:2017-11-21
  • Insights into a Hole Transfer Mechanism between Glucose Oxidase and a p-type Organic Semiconductor
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-21
    Gintautas Bagdžiūnas, Šarūnas Žukauskas, Arūnas Ramanavičius

    This manuscript describes a bioelectrochemical application of a new class of electrochemically generated hole-transporting (p-type) polymeric semiconductors (HTPS), which are based on carbazole core and the oxiran and thiiran reactive groups. Electrode based on transparent layer of indium tin oxide was electrochemically modified with a layer of HTPS and a monolayer of covalently immobilized glucose oxidase (GOx). The HTPS/GOx-based electrode was investigated for an evaluation of direct hole-transfer between the enzyme and electrode at a bio-electrochemically relevant potential via HTPS layer. The broad linear relationship between the peak-current density and glucose concentration from 2 to 15 mM and high stability of ITO/poly-CzS/GOx-electrode was observed. Moreover, it was determined that charge transfer rate constants are reliable for the establishment of advanced electron transfer between enzyme and electrode for the application of this HTPS/GOx-based electrode in long-lived biofuel cells and amperometric biosensors.

    更新日期:2017-11-21
  • A novel electrochemical sensor based on Cu@Ni/MWCNTs nanocomposite for simultaneous determination of guanine and adenine
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-20
    Dongyang Wang, Bintong Huang, Jie Liu, Xia Guo, Guzailinur Abudukeyoumu, Yang Zhang, Bang-Ce Ye, Yingchun Li

    A novel electrochemical sensing platform based on combination of multi-walled carbon nanotubes and copper-nickel hybrid nanoparticles (Cu@Ni/MWCNTs) was developed for simultaneous detection of guanine (G) and adenine (A). The Ni/MWCNTs and Cu@Ni/MWCNTs nanocomposites were characterized by transmission electron microscopy (TEM) and energy-dispersive X-ray spectroscopy (EDS). The electrochemical behaviors of G and A on the modified electrode were explored by cyclic voltammetry (CV) and differential pulse voltammetry (DPV) in phosphate buffer with pH 3.0. Under the optimal conditions, electrical signals were linear over the concentration ranges from 5.0 to 180 μM and 8.0 to 150 μM for simultaneous determination G and A with the detection limit as low as 0.35 μM and 0.56 μM (S/N=3), respectively. Furthermore, linear concentration ranges in individual determination are 1.0~180 μM and 2.0~150 μM with detection limits of 0.17 μM and 0.33 μM (S/N=3) for G and A, respectively. The sensor was successfully used to quantify G and A in real samples. The Cu@Ni/MWCNTs composite presented here can serve as a promising candidate for developing electrochemical sensor devices and plays an important role in widespread fields.

    更新日期:2017-11-20
  • Paper-based diagnostics in the antigen-depletion regime: high-density immobilization of rcSso7d-cellulose-binding domain fusion proteins for efficient target capture
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-20
    Eric A. Miller, Subha Baniya, Daniel Osorio, Yara Jabbour Al Maalouf, Hadley D. Sikes

    In this work, we report the development of a general strategy for enhancing the efficiency of target capture in immunoassays, using a bifunctional fusion protein construct which incorporates a substrate-anchoring moiety for the high-abundance immobilization of an antigen-binding domain. This approach was informed by the development of a pseudo first-order rate constant model, and tested in a paper-based assay format using a fusion construct consisting of an rcSso7d binding module and a cellulose-binding domain. These rcSso7d-CBD fusion proteins were solubly expressed and purified from bacteria in high molar yields, and enable oriented, high-density adsorption of the rcSso7d binding species to unmodified cellulose within a 30-second incubation period. These findings were validated using two distinct, antigen-specific rcSso7d variants, which were isolated from a yeast surface display library via flow cytometry. Up to 1.6 micromoles of rcSso7d-CBD was found to adsorb per gram of cellulose, yielding a local binder concentration of up to 760 μM within the resulting active material. At this molar abundance, target antigens are captured from solution with nearly 100% efficiency, maximizing the attainable sensitivity for any given diagnostic system.

    更新日期:2017-11-20
  • Porous silicon based photoluminescence immunosensor for rapid and highly-sensitive detection of Ochratoxin A
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-17
    Valerii Myndrul, Roman Viter, Maryna Savchuk, Nelya Shpyrka, Donats Erts, Daniel Jevdokimovs, Viesturs Silamiķelis, Valentyn Smyntyna, Arunas Ramanavicius, Igor Iatsunskyi

    A rapid and low cost photoluminescence (PL) immunosensor for the determination of low concentrations of Ochratoxin A (OTA) has been developed. This immunosensor was based on porous silicon (PSi) and modified by antibodies against OTA (anti-OTA). PSi layer was fabricated by metal-assisted chemical etching (MACE) procedure. Main structural parameters (pore size, layer thickness, morphology and nanograins size) and composition of PSi were investigated by means of X-Ray diffraction (XRD), scanning electron microscopy (SEM) and Raman spectroscopy. PL-spectroscopy of PSi was performed at room temperature and showed a wide emission band centered at 680±20 nm. Protein A was covalently immobilized on the surface of PSi, which in next steps was modified by anti-OTA and BSA in this way a anti-OTA/Protein-A/PSi structure sensitive towards OTA was designed. The anti-OTA/Protein-A/PSi-based immunosensors were tested in a wide range of OTA concentrations from 0.001 upto 100 ng/ml. Interaction of OTA with anti-OTA/Protein-A/PSi surface resulted in the quenching of photoluminescence in comparison to bare PSi. The limit of detection (LOD) and the sensitivity range of anti-OTA/Protein-A/PSi immunosensors were estimated. Association constant and Gibbs free energy for the interaction of anti-OTA/Protein-A/PSi with OTA were calculated and analyzed using the interaction isotherms. Response time of the anti-OTA/Protein-A/PSi-based immunosensor toward OTA was in the range of 500–700 s. These findings are very promising for the development of highly sensitive, and potentially portable immunosensors suitable for fast determination of OTA in food and beverages.

    更新日期:2017-11-20
  • Selective isolation and noninvasive analysis of circulating cancer stem cells through Raman imaging
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-16
    Hyeon-Yeol Cho, Khaled Hossain, Jin-Ho Lee, Jiyou Han, Hun Joo Lee, Kyeong-Jun Kim, Jong-Hoon Kim, Ki-Bum Lee, Jeong-Woo Choi

    Circulating cancer stem cells (CCSCs), a rare circulating tumor cell (CTC) type, recently arose as a useful resource for monitoring and characterizing both cancers and their metastatic derivatives. However, due to the scarcity of CCSCs among hematologic cells in the blood and the complexity of the phenotype confirmation process, CCSC research can be extremely challenging. Hence, we report a nanoparticle-mediated Raman imaging method for CCSC characterization which profiles CCSCs based on their surface marker expression phenotypes. We have developed an integrated combinatorial Raman-Active Nanoprobe (RAN) system combined with a microfluidic chip to successfully process complete blood samples. CCSCs and CTCs were detected (90% efficiency) and classified in accordance with their respective surface marker expression via completely distinct Raman signals of RANs. Selectively isolated CCSCs (93% accuracy) were employed for both in vitro and in vivo tumor phenotyping to identify the tumorigenicity of the CCSCs. We utilized our new method to predict metastasis by screening blood samples from xenograft models, showing that upon CCSC detection, all subjects exhibited liver metastasis. Having highly efficient detection and noninvasive isolation capabilities, we have demonstrated that our RAN-based Raman imaging method will be valuable for predicting cancer metastasis and relapse via CCSC detection. Moreover, the exclusion of peak overlapping in CCSC analysis with our Raman imaging method will allow to expand the RAN families for various cancer types, therefore, increasing therapeutic efficacy by providing detailed molecular features of tumor subtypes.

    更新日期:2017-11-20
  • Porous Ni0.1Mn0.9O1.45 microellipsoids as high-performance anode electrocatalyst for microbial fuel cells
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-16
    Lizhen Zeng, Wenguang Zhang, Pan Xia, Wenqiang Tu, Changchun Ye, Miao He

    A novel bi-component composite of porous self-assembled micro-/nanostructured Ni0.1Mn0.9O1.45 microellipsoids as high-performance anode electrocatalyst for microbial fuel cells (MFCs) is successfully synthesized via a simple coprecipitation reaction in microemulsion and calcination method in air atmosphere. The morphology and structural characterization indicate that the as-fabricated Ni0.1Mn0.9O1.45 product is consist of Mn2O3 and NiMn2O4 (n(Mn2O3): n(NiMn2O4) = 0.35: 0.1) and has a porous microellipsoidal morphology. The microellipsoids are compose of numerous layered micro-/nanostructured blocks and the special porous microellipsoids structure of Ni0.1Mn0.9O1.45 offers a large specific surface area for bacteria adhesion. The porous Ni0.1Mn0.9O1.45 microellipsoids as anode electrocatalyst for MFCs exhibits excellent electrocatalytic activity to promote the extracellular electron transfer (EET) between the anode and bacteria, hence improves the performance of MFC. The MFC equipped with Ni0.1Mn0.9O1.45/CF anode achieves a maximum power density of 1.39 ± 0.02 W m−2, is significantly higher than that of commercial carbon felt anode. This work proposes a new method for the synthesis of high-performance and environmentally friendly anode electrocatalyst for MFCs.

    更新日期:2017-11-17
  • A SELF-POWERED BIOSENSING DEVICE WITH AN INTEGRATED HYBRID BIOFUEL CELL FOR INTERMITTENT MONITORING OF ANALYTES
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-16
    Dominika Majdecka, Sylwia Draminska, Dariusz Janusek, Paweł Krysinski, Renata Bilewicz

    In this work, we propose an integrated self-powered sensing system, driven by a hybrid biofuel cell (HBFC) with carbon paper discs coated with multiwalled carbon nanotubes. The sensing system has a biocathode made from laccase or bilirubin oxidase, and the anode is made from a zinc plate. The system includes a dedicated custom-built electronic control unit for the detection of oxygen and catechol analytes, which are central to medical and environmental applications. Both the HBFC and sensors, operate in a mediatorless direct electron transfer mode. The measured characteristics of the HBFC with externally applied resistance included the power-time dependencies under flow cell conditions, the sensors performance (evaluated by cyclic voltammetry), and chronoamperometry. The HBFC is integrated with analytical devices and operating in a pulse mode form long-run monitoring experiments. The HBFC generated sufficient power for wireless data transmission to a local computer.

    更新日期:2017-11-17
  • A novel 3D bioprinted flexible and biocompatible hydrogel bioelectronic platform
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-16
    Shweta Agarwala, Jia Min Lee, Wei Long Ng, Michael Layani, Wai Yee Yeong, Shlomo Magdassi

    Bioelectronics platforms are gaining widespread attention as they provide a template to study the interactions between biological species and electronics. Decoding the effect of the electrical signals on the cells and tissues holds the promise for treating the malignant tissue growth, regenerating organs and engineering new-age medical devices. This work is a step forward in this direction, where bio- and electronic materials co-exist on one platform without any need for post processing. We fabricate a freestanding and flexible hydrogel based platform using 3D bioprinting. The fabrication process is simple, easy and provides a flexible route to print materials with preferred shapes, size and spatial orientation. Through the design of interdigitated electrodes and heating coil, the platform can be tailored to print various circuits for different functionalities. The biocompatibility of the printed platform is tested using C2C12 murine myoblasts cell line. Furthermore, normal human dermal fibroblasts (primary cells) are also seeded on the platform to ascertain the compatibility.

    更新日期:2017-11-17
  • Ultrasensitive detection of prostate specific antigen by electrochemical aptasensor using enzyme-free recycling amplification via target-induced catalytic hairpin assembly
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-15
    Juncai Zhao, Zhanfang Ma

    Based on the target-induced catalytic hairpin assembly and bimetallic catalyst, the enzyme-free recycling amplification strategy for sensitive detection of prostate specific antigen (PSA) has been designed. The aptamer and its complementary DNA (C-apt) are modified on the magnetic particles. The aptamer-PSA binding event can release the C-apt that triggers the catalytic assembly between hairpin capture DNA and hairpin help DNA. Then the catalytic hairpin assembly leads to cyclic reuse the C-apt and the generation of many opened hairpin capture DNA, which can associate with the prepared Au/Pt-polymethylene blue (PMB) probes to yield electrochemical signal. Meanwhile, the Au/Pt-PMB probes exhibit excellent electrocatalytic ability for H2O2 to magnify the response current. The designed sensor possesses a wide dynamic range of 10 fg mL−1 to 100 ng mL−1 and ultra-low detection limit of 2.3 fg mL−1. The present method has good performance in real serum sample analysis. This strategy is promising to be extended to provide a highly sensitive platform for various target analytes.

    更新日期:2017-11-16
  • Target-catalyzed hairpin assembly and metal-organic frameworks mediated nonenzymatic co-reaction for multiple signal amplification detection of miR-122 in human serum
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-15
    Yuliang Li, Chao Yu, Bo Yang, Zhirui Liu, Peiyuan Xia, Qian Wang

    Herein, a new type of multifunctional iron based metal-organic frameworks (PdNPs@Fe-MOFs) has been synthesized by assembly palladium nanoparticles on the surface of Fe-MIL-88NH2 MOFs microcrystals, and first applied in electrochemical biosensor for ultrasensitive detection of microRNA-122 (miR-122, a biomarker of drug-induced liver injury). The nanohybrids have not only been utilized as ideal nanocarriers for immobilization of signal probes, but also used as redox probes and electrocatalysts. In this biosensor, two hairpin probes were designed as capture probes and signal probes, respectively. The nanohybrids conjugated with streptavidin and biotinylated signal probes were used as the tracer labels, target miR-122 was sandwiched between the tracer labels and thiol-terminated capture probes inserted in MCH monolayer on the gold nanoparticles-functionalized nitrogen-doped graphene sheets (AuNPs@N-G) modified electrode. Based on target-catalyzed hairpin assembly, target miR-122 could trigger the hybridization of capture probes and signal probes to further be released to initiate the next reaction process resulted in numerous tracer indicators anchored onto the sensing interfaces. Thus, the detection signal could be dramatically enhanced towards the electrocatalytic oxidation of 3,3’,5,5’-tetramethylbenzidine in the presence of H2O2 owing to the intrinsic and intriguing peroxidase-like activity of the nanohybrids. With the assist of target-catalyzed hairpin assembly and PdNPs@Fe-MOFs mimetic co-reaction for signal amplification, a wide detection range from 0.01 fM to 10 pM was achieved with a low detection limit of 0.003 fM (S/N=3). Furthermore, the proposed biosensor exhibited excellent specificity and recovery in spiked serum samples, and was successfully used for detecting miR-122 in real biological samples, which provided a rapid and efficient method for detecting drug-induced liver injury at an early stage.

    更新日期:2017-11-16
  • Poly (dopamine quinone-chromium (III) complex) microspheres as new modifier for simultaneous determination of phenolic compounds
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-12
    M. Shahbakhsh, M. Noroozifar

    In this study, poly(dopamine-quinone chromium (III))-microspheres (PDQCM) were used for the modification of graphite paste electrode (GPE) for simultaneous voltammetric determination of 4-Amino Phenol (AP), Phenol (Ph) and 4-Nitro Phenol (NP). The PDQCM and the GPE modified with PDQCM were characterized by field emission scanning electron microscopy, Energy-dispersive X-ray spectroscopy, Fourier transform infrared spectroscopy, electrochemical impedance spectroscopy and electrochemistry methods such as cyclic voltammetry (CV) and differential pulse voltammetry (DPV). The proposed modified electrode exhibits high electrocatalytic activity toward electrooxidation of AP, Ph, and NP to three well-separated peaks in the potential range from 0.2 to 1.3 V using CV and DPV methods in phosphate buffer solution with pH 2.0. Under the optimum conditions, detection limits of 0.5, 0.6 and 0.8 µM were obtained for AP, Ph, and NP, respectively. Moreover, GPE/PDQCM was successfully used for simultaneous determination of AP, Ph, and NP in tap and river waters.

    更新日期:2017-11-13
  • A novel screen-printed mast cell-based electrochemical sensor for detecting spoilage bacterial quorum signaling molecules (N-acyl-homoserine-lactones) in freshwater fish
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-11
    Donglei Jiang, Yan Liu, Hui Jiang, Shengqi Rao, Wu Fang, Mangang Wu, Limin Yuan, Weiming Fang

    A novel screen-printed cell-based electrochemical sensor was developed to assess bacterial quorum signaling molecules, N-acylhomoserine lactones (AHLs). Screen-printed carbon electrode (SPCE), which possesses excellent properties such as low-cost, disposable and energy-efficient, was modified with multi-walled carbon nanotubes (MWNTs) to improve electrochemical signals and enhance the sensitivity. Rat basophilic leukemia (RBL-2H3) mast cells encapsulated in alginate/graphene oxide (NaAgl/GO) hydrogel were immobilized on the MWNTs/SPCE to serve as recognition element. Electrochemical impedance spectroscopy (EIS) was employed to record the cell impedance signal as-influenced by Pseudomonas aeruginosa quorum-sensing molecule, N−3-oxododecanoyl homoserine lactone (3OC12-HSL). Experimental results show that 3OC12-HSL caused a significant decrease in cell viability in a dose dependent manner. The EIS value decreased with concentrations of 3OC12-HSL in the range of 0.1 to 1 μM, and the detection limit for 3OC12-HSL was calculated to be 0.094 μM. These results were confirmed via cell viability, SEM, TEM analysis. Next, the sensor was successfully applied to monitoring the production of AHLs by spoilage bacteria in three different freshwater fish juice samples which efficiently proved the practicability of this cell based method. Therefore, the proposed cell sensor may serve as an innovative and effective approach to the measurement of quorum signaling molecule and thus provides a new avenue for real-time monitoring the spoilage bacteria in freshwater fish production.

    更新日期:2017-11-13
  • A novel label-free electrochemical immunosensor for ultra-sensitively detecting prostate specific antigen based on the enhanced catalytic currents of oxygen reduction catalyzed by core-shell Au@Pt nanocrystals
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-11
    Rui Wang, Ai-Jun Wang, Wei-Dong Liu, Pei-Xin Yuan, Yadong Xue, Xiliang Luo, Jiu-Ju Feng

    Herein, bimetallic core-shell Au@Pt nanocrystals (Au@Pt NCs) were prepared by a simple one-pot aqueous method using 2-pyrrolidone-5-carboxylic acid sodium salt (PCA-Na) as a new and green growth-directing agent. The obtained architectures displayed excellent catalytic activity towards oxygen reduction reaction (ORR) compared with commercial Pt/C catalyst. A novel immunosensor was constructed via assembly prostate specific antibody on the surface of Au@Pt NCs. It was found that the ORR currents were significantly suppressed due to the specific antigen-antibody reaction. The ultra-sensitive determination of prostate specific antigen (PSA) was realized on account of the immunocomplex impeding the redox probe accessible to the electrode. The immunosensor exhibited good analytical performance for the assay of PSA with the wide linear range of 0.1 ~ 50 ng mL–1 and low detection limit of 0.018 ng mL–1 (S/N = 3), coupled with the improved stability, reproducibility and selectivity.

    更新日期:2017-11-13
  • Cu@Pd core-shell nanostructures for highly sensitive and selective amperometric analysis of histamine
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-11
    Gajjala Rajendra Kumar Reddy, Palathedath Suresh kumar

    We demonstrate a facile and rapid methodology for preparation of Cu@Pd core-shell nanostructures on a cost-effective pencil graphite substrate. Galvanic replacement reaction was carried out for palladium modification on template electrodeposited copper nanostructures on pencil graphite substrate. The nanostructures are shown to be very stable with excellent electrocatalytic activities. Under optimised conditions, they could be used for histamine sensing at a very low oxidation potential of +0.55 V vs. Ag/AgCl. The low oxidation potential enabled sensitive and selective analysis of histamine using chronoamperometry without any interference from oxygen evolution reactions. We have demonstrated that the sensor shows excellent selectivity towards histamine even in the presence of many of the common interfering biogenic amines. The sensor exhibited a sensitivity of 0.082 μ μ A/μ μ M/cm 2 2 with a a limit of detection as low as 3.2 ± ± 0.1 nM. The oxidation potential and limit of detection obtained using this sensor are much superior to the results reported so far in the literature. Practical feasibility of the developed sensor was manifested by histamine analysis in canned tuna fish samples, where the chronoamperometric estimation was also validated by conventional HPLC analysis.

    更新日期:2017-11-13
  • Graphene and Au NPs co-mediated enzymatic silver deposition for the ultrasensitive electrochemical detection of cholesterol
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-11
    Yong Huang, Jie Tan, Lijie Cui, Zhide Zhou, Zhenghua Zhang, Rong Zheng, Yewei Xue, Mengxin Zhang, Shanshan Li, Nixuan Zhu, Jintao Liang, Guiyin Li, Sufang Zhou, Yongxiang Zhao

    Cholesterol is an essential ingredient in mammals, and serum cholesterol is a major component of atherosclerotic plaques. The level of cholesterol in human serum has become an important index for clinical diagnosis and prevention of cardiovascular disease. In this paper, a simple and ultrasensitive cholesterol biosensor based on graphene oxide (GO) and gold nanoparticles (Au NPs) co-mediated enzymatic silver deposition was designed by immobilizing cholesterol oxidase (CHOD), cholesterol esterase (CHER) and GO onto the surface of Au NPs modified screen-printed carbon electrode (SPE). Under the synergistic effect of CHER, CHOD and GO, the cholesterol was hydrolyzed to generate hydrogen peroxide, which can reduce the silver (Ag) ions in the solution to metallic Ag which deposited on the surface of Au NPs modified SPE. The ultrasensitive detection of cholesterol was achieved by anodic stripping voltammetry measurement of the enzymatically deposited Ag. Under optimal conditions, the anodic stripping peak current of Ag increased with the increasing cholesterol concentration in the range from 0.01 μg/mL to 5000 μg/mL with a limit of detection of 0.001 μg/mL (S/N=3). In addition, the ultrasensitive cholesterol biosensor exhibited higher specificity, acceptable reproducibility and excellent recoveries for cholesterol detection.

    更新日期:2017-11-11
  • Detection of early stage prostate cancer by using a simple carbon nanotube@paper biosensor
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-10
    Sungkyung Ji, Myeongsoon Lee, Don Kim

    This study is an investigation for an inexpensive, simple and sensitive biosensor to detect prostate cancer using bioactivated-multi wall carbon nanotubes (MWCNTs, diameter of 20 nm, length of 5 μm) and a micro-pore filter paper (pore size of 0.45 μm). For the immunoassay of prostate specific antigen (PSA), which is a biomarker of prostate cancer, MWCNTs were activated with PSA antibody (monoclonal antibody of the prostate specific antigen) by using N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysulfosuccinimide sodium salt (NHSS). The activated MWCNTs were deposited on the micro-pore filter paper to use as a biosensor. The prepared biosensor can assay from 0 to 500 ng/mL of PSA level within 2 h with the detection limit of 1.18 ng/mL by the measurement of resistance change. The resistance change was caused by site selective interaction between PSA and PSA-antigen with an inexpensive bench top digital multimeter (5 1/2 digits). The detection range and sensitivity of the prepared sensor are good enough to diagnose the early stage of prostate cancer (> 4 ng/mL of PSA). This paper-based biosensor is about 20 times cheaper (fabricated biosensor price: 2.4 $) and over 10 times faster than enzyme-linked immunosorbent assay (ELISA), which is a general method for the detection of a specific protein in the modernized hospitals. Furthermore, the maximum detection limit is about 50 times higher than ELISA.

    更新日期:2017-11-11
  • Switchable electrochemiluminescence aptasensor coupled with resonance energy transfer for selective attomolar detection of Hg2+ via CdTe@CdS/dendrimer probe and Au nanoparticle quencher
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-10
    Bahareh Babamiri, Abdollah Salimi, Rahman Hallaj

    In the present study, an ultrasensitive electrochemiluminescence (ECL) aptasensing assay for selective detection of Hg2+ was designed. In this electrochemiluminescence resonance energy transfer (ECL-RET) approach, Fe3O4@SiO2/dendrimers/QDs exhibited amplified ECL emissions (switch “on” state) and with the hybridization between T-rich ssDNA(S1) immobilized on the Fe3O4@SiO2/dendrimers/QDs and AuNPs modified with complementary aptamer (AuNPs-S2), the ECL of QDs nanocomposites was efficiently quenched (switch “off” state). In the presence of Hg2+ ions, formation of strong and stable T-Hg2+-T complex led to the release of the AuNPs-S2 from double-stranded DNA(dsDNA) and the recovery of the ECL signal of QDs (second signal switch “on” state). Under optimal conditions, Hg2+ can be detected in a wide linear range from 20 aM to 2 µM with a very low detection limit of 2 aM. The proposed ECL aptasensor showed high selectivity for Hg2+ determination compared to other environmentally relevant metal ions at concentration ratio more than 1000 times. The aptasensor was used for detection Hg2+ ions from samples of tap waters, carp and saltwater fishes with satisfactory results. The aptasensor exhibited high sensitivity, wide linear response (11 orders of magnitude), excellent reproducibility and stability. The proposed aptasensor will be a promising candidate for facile and rapid determination of Hg2+in environmental and fishery samples.

    更新日期:2017-11-10
  • Effect of Brownian motion on reduced agglomeration of nanostructured metal oxide towards development of efficient cancer biosensor
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-10
    Suveen Kumar, Ashish, Saurabh Kumar, Shine Augustine, Santosh Yadav, Birendra Kumar Yadav, Rishi Pal Chauhan, Ajay Kumar Dewan, Bansi Dhar Malhotra

    We report results of the studies relating to fabrication of nanostructured metal oxide (NMO) based cancer biosensor. With the help of 2D electroactive reduced graphene oxide (RGO), we successfully inhibited the Brownian motion of NMO that led to reduced agglomeration of NMO. The nanostructured hafnium oxide (nHfO2) was used as a model NMO. The reduced agglomeration of nHfO2 was achieved through controlled hydrothermal synthesis and investigated via nanoparticles tracking analysis (NTA). X-ray diffraction (XRD), scanning electron microscopy (SEM) and transmission electron microscope (TEM) techniques were used for phase identification as well as morphological analysis of the synthesized nanohybrid (nHfO2@RGO) material. The 3-aminopropyl triethoxysilane (APTES) was used for the functionalization of nHfO2@RGO and electrophoretic deposition (EPD) technique was used for its deposition onto ITO coated glass electrode. Further, antibodies of cancer biomarker (anti-CYFRA-21-1) were immobilized via EDC-NHS chemistry and Bovine serum albumin (BSA) was used for blocking of the non-specific binding sites. The electrochemical response studies of fabricated immunoelectrode (BSA/anti-CYFRA-21-1/APTES/nHfO2@RGO/ITO) revealed higher sensitivity (18.24µAmLng-1), wide linear detection range (0 to 30ngmL-1), with remarkable lower detection limit (0.16ngmL-1). The obtained results showed good agreement with the concentration of CYFRA-21-1 obtained through enzyme linked immunosorbent assay (ELISA) in saliva samples of oral cancer patients.

    更新日期:2017-11-10
  • Ultrasensitive electrochemical immunosensor for quantitative detection of HBeAg using Au@Pd/MoS2@MWCNTs nanocomposite as enzyme-mimetic labels
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-09
    Zengqiang Gao, Yueyun Li, Xiaobo Zhang, Jinhui Feng, Ling Kong, Ping Wang, Zhiwei Chen, Yunhui Dong, Qin Wei

    A sensitive sandwich-type electrochemical immunosensor for the detection of hepatitis B e antigen (HBeAg) was successfully developed based on the gold@palladium nanoparticles (Au@Pd NPs) loaded by molybdenum disulfide functionalized multiwalled carbon nanotubes (Au@Pd/MoS2@MWCNTs). The resultant nanocomposites not only possessed high specific surface area and good biocompatibility, but also exhibited excellent electro-catalytical property. Au NPs functionalized porous graphene oxide (p-GO@Au) were used as sensing platforms and primary antibodies carriers, which can accelerate the electron transfer and improve the load capacity of primary antibodies (Ab1), improving the sensitivity of the immunosensor. Under optimal conditions, the designed immunosensor could detect target HBeAg concentration in the range from 0.1 pg/mL to 500 pg/mL, with a low detection limit of 26 fg/mL (S/N=3) for HBeAg. Additionally, the designed immunosensor showed excellent specificity, good reproducibility and acceptable stability. The satisfactory results in analysis of human serum samples indicated that it had potential application in clinical monitoring of tumor markers.

    更新日期:2017-11-10
  • Pyridoxamine driven selective turn-off detection of picric acid using glutathione stabilized fluorescent copper nanoclusters and its applications with chemically modified cellulose strips
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-09
    Ravi Patel, Shilpa Bothra, Rajender Kumar, Guido Crisponi, Suban K Sahoo

    The present work reports the interaction of various vitamin B6 cofactors with the red emitting glutathione stabilized copper nanoclusters (GSH-CuNCs). Addition of pyridoxamine (PM) resulted a new turn-on band at 410 nm due to the possible adsorption over the surface of GSH-CuNCs. The nano-assembly PM-GSH-CuNCs was applied for the selective detection of nitro-aromatic compounds. Upon addition of picric acid (PA), the fluorescence of PM-GSH-CuNCs was selectively quenched at 410 nm and ~625 nm among the other tested nitro-aromatic compounds. With a linearity range from 9.9 μM to 43 μM, the concentration of PA can be detected down to 2.74 μM. The high selectivity exhibited by the nano-assembly allows to detect PA in real samples like tap water, river water and matchstick. Advantageously, the nano-assembly PM-GSH-CuNCs was chemically adsorbed over the cellulosic strips and applied for the naked-eye detection of PA down to 1 μM.

    更新日期:2017-11-10
  • Target-induced proximity ligation triggers recombinase polymerase amplification and transcription-mediated amplification to detect tumor-derived exosomes in nasopharyngeal carcinoma with high sensitivity
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-09
    Wanli Liu, Jianpei Li, Yixian Wu, Shan Xing, Yanzhen Lai, Ge Zhang

    Tumor-derived exosomes (TEXs) are extracellular vesicles that are continuously released into the blood by tumor cells and carry specific surface markers of the original tumor cells. Substantial evidence has implicated TEXs as attractive diagnostic markers for cancer. However, the detection of TEXs in blood at an early tumor stage is challenging due to their very low concentration. Here, we established a method called PLA-RPA-TMA assay that allows TEXs to be detected with high sensitivity and specificity. Based on two proximity ligation assay (PLA) probes that recognize a biomarker on a TEX, we generated a unique surrogate DNA signal for the specific biomarker, which was synchronously amplified twice by recombinase polymerase amplification (RPA) coupled with transcription-mediated amplification (TMA), and then the products of the RPA-TMA reaction were quantitatively detected using a gold nanoparticle-based colorimetric assay. We established proof-of-concept evidence for this approach using TEXs from nasopharyngeal carcinoma (NPC) cells, with a detection limit of 102 particles/mL, and reported the measurement of plasma Epstein-Barr virus latent membrane protein 1 (LPM1)-positive (LMP1+, accuracy: 0.956) and epidermal growth factor receptor (EGFR)-positive (EGFR+, accuracy: 0.906) TEXs as potent early diagnostic biomarkers for NPC.

    更新日期:2017-11-10
  • Nucleic acid aptamer-based methods for diagnosis of Infections
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-08
    Ki Soo Park

    Infectious diseases are a serious global problem, which not only take an enormous human toll but also incur tremendous economic losses. In combating infectious diseases, rapid and accurate diagnostic tests are required for pathogen identification at the point of care (POC). In this review, investigations of diagnostic strategies for infectious diseases that are based on aptamers, especially nucleic acid aptamers, oligonucleotides that have high affinities and specificities toward their targets, are described. Owing to their unique features including low cost of production, easy chemical modification, high chemical stability, reproducibility, and low levels of immunogenicity and toxicity, aptamers have been widely utilized as bio-recognition elements (bio-receptors) for the development of infection diagnostic systems. In this review, we discuss nucleic acid aptamer-based methods that have been developed for diagnosis of infections using a format that organizes discussion according to the target pathogenic analytes including toxins or proteins, whole cells and nucleic acids. Also included is, a summary of recent advances made in the sensitive detection of pathogenic bacteria utilizing the isothermal nucleic acid amplification method. Lastly, a nucleic acid aptamer-based POC system is described and future directions of studies in this area are discussed.

    更新日期:2017-11-10
  • ELECTROCHEMICAL IMMUNOSENSORS – A POWERFUL TOOL FOR ANALYTICAL APPLICATIONS
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-08
    Fabiana S. Felix, Lúcio Angnes

    Immunosensors are biosensors based on interactions between an antibody and antigen on a transducer surface. Either antibody or antigen can be the species immobilized on the transducer to detect antigen or antibody, respectively. Because of the strong binding forces between these biomolecules, immunosensors present high selectivity and very high sensitivity, making them very attractive for many applications in different science fields. Electrochemical immunosensors explore measurements of an electrical signal produced on an electrochemical transductor. This signal can be voltammetric, potentiometric, conductometric or impedimetric. Immunosensors utilizing electrochemical detection have been explored in several analyses since they are specific, simple, portable, and generally disposable and can carry out in situ or automated detection. This review addresses the potential of immunosensors destined for application in food and environmental analysis, and cancer biomarker diagnosis. Emphasis is given to the approaches that have been used for construction of electrochemical immunosensors. Additionally, the fundamentals of immunosensors, technology of transducers and nanomaterials and a general overview of the possible applications of electrochemical immunosensors to the food, environmental and diseases analysis fields are described.

    更新日期:2017-11-10
  • Ratiometric Electrochemical Assay for Sensitive Detecting MicroRNA Based on Dual-amplification Mechanism of Duplex-specific Nuclease and Hybridization Chain Reaction
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-08
    Yan-Hong Yuan, Bao-Zhu Chi, Shao-Hua Wen, Ru-Ping Liang, Zhi-Mei Li, Jian-Ding Qiu

    We propose a ratiometric electrochemical assay for detecting microRNA (miRNA) on the basis of dual-amplification mechanism by using distinguishable electrochemical signals from thionine (Thi) and ferrocene (Fc). The thiol-modified and ferrocene-labeled hairpin capture probes (CP) are first immobilized on an Au electrode via Au-S reaction. The target miRNA hybridizes with CP and unfolding the hairpin structure of CP to form miRNA-DNA duplexes. Then, kamchatka crab duplex specific nuclease (DSN) specifically cleaves the DNA in miRNA-DNA duplexes, leading to the release of miRNA and another cleaves cycle, meanwhile, numerous Fc leaves away from the electrode surface and leads to the signal-off of Fc. The residual fragment on electrode surface acts as a HCR primer to form dsDNA polymers through in situ HCR with the presence of the primer and two probes (HDNA and HDNA’), resulting in the capture of numerous DNA/Au NPs/Thi and the signal-on of Thi. The dual-amplification mechanism significantly amplifies the decrease of Fc signal and the increase of Thi signal for ratiometric readout (IThi/IFc), thus providing a sensitive method for the selective detection of miR-141 with a detection limit down to 11 aM. The dual-signal ratiometric outputs have an intrinsic self-calibration to the effects from system, which is promising to be applied in biosensing and clinical diagnosis.

    更新日期:2017-11-10
  • Ultrahigh sensitive enhanced-electrochemiluminescence detection of cancer biomarkers using silica NPs/graphene oxide; a comparative study
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-08
    Hamid Kooshki, Jamal Rashidiani, Mehdi Kamali, Hamid Sedighian, Mostafa Akbariqomi, Mansour Mansouri

    The increasing progress in using nano-biomaterials for medical purposes has opened new horizons toward researchers around the globe. To investigate the presence of these nanomaterials and the impacts they might have, a comparative enhanced-electrochemiluminescence immunosensing study has been designed. The effects of utilizing graphene oxide, silica, and gold nanoparticles in cancer diagnosis were evaluated during the quantification of two major cancer biomarkers (CEA and AFP) in different approaches. In other words, first and second approaches were designed to employ nanomaterials while third and fourth approaches were developed in absence of those. Accordingly, resulted LODs experienced dramatic amplification when nano-biomaterials were included in the immunosensor modification (for AFP: 1st and 3rd approaches: 1.36 fg/ml in comparison with 0.39 ng/ml, and for CEA: 2nd and 4th approaches: 1.90 fg/ml versus 0.46 ng/ml, respectively). Correspondingly, capability of nano-biomaterials for developing highly sensitive and more efficient immunosensors was validated through selectivity, stability, reproducibility, and feasibility examinations.

    更新日期:2017-11-10
  • High-purity Capture of CTCs Based on Micro-beads Enhanced Isolation by Size of Epithelial Tumor Cells (ISET) Method
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-07
    Na Sun, Xinpan Li, Zhili Wang, Yuzhi Li, Renjun Pei

    In this paper, we develop a low-cost size-based microfluidic chip using conventional polycarbonate membrane to isolate CTCs from blood, and propose a strategy to increase the capture efficiency before cell filtration by a size enlargement method utilizing modified microbeads specifically binding to CTCs. Up to 91% of target cells were isolated from whole blood samples using our microfluidic capture system at a flow rate of 1 mL/min. Moreover, a WBC depletion process is introduced which greatly decreases the WBC retaining on the filter membrane. The tests of immunofluorescence analysis of cells captured on the membrane were performed, which demonstrates that the device could provide a dependable CTC identification and CTC count in whole blood samples. Finally, the device was further validated in the detection of CTCs from blood samples of cancer patients, and it indicates a promising capability to detect CTC response to treatment.

    更新日期:2017-11-10
  • A facile way to fabricate manganese phosphate self-assembled carbon networks as efficient electrochemical catalysts for real-time monitoring of superoxide anions released from HepG2 cells
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-06
    Xuan Cai, Libo Shi, Wenqian Sun, Hongli Zhao, Hong Li, Haiyan He, Minbo Lan

    Quantification of superoxide anions (O2•−) is significant in the monitoring of many serious diseases and the design of enzyme-mimic catalysts plays the main role in the development of non-enzymatic O2•− sensors. Herein, we proposed a facile self-assembly process to synthesize manganese phosphate modified carbon networks using three kinds of widely-used carbon materials (MWCNTs, NGS and GO) as pillar connectors. Characterizations demonstrate that manganese phosphate is widely dispersed inside and on the surface of carbon networks without visible morphology. Meanwhile, all three kinds of synthesized catalysts were successfully immobilized on the screen-printed carbon electrodes to evaluate the electrochemical performance of fabricated sensors. The results indicate that sensors based on Mnx(PO4)y modified MWCNTs exhibit high sensitivity with an extremely low detection limit of 0.127 μM (S/N = 3) and a wide liner range of 0–1.817 mM (R2 = 0.998). We further employed the recommended sensors in the real-time monitoring of HepG2 cells released O2•− under the stimulating of Zymosan (20 mg/mL). Noticeably, the proposed sensors exhibit not only sensitive response but also stable current steps upon different addition of Zymosan. The calculated concentrations of cell-released O2•− vary from 6.772 to 24.652 pM cell-1 for the Zymosan ammount used in this work. The established novel sensors display low background current and signal noises, thus holding unique advantages in the trace analysis of O2•− in biological samples and in vivo environment.

    更新日期:2017-11-10
  • Pyrrolidinyl PNA polypyrrole/silver nanofoam electrode as a novel label-free electrochemical miRNA-21 biosensor
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-06
    Tawatchai Kangkamano, Apon Numnuam, Warakorn Limbut, Proespichaya Kanatharana, Tirayut Vilaivan, Panote Thavarungkul

    A label-free electrochemical miRNA biosensor was developed based on a pyrrolidinyl peptide nucleic acid (acpcPNA)/polypyrrole (PPy)/silver nanofoam (AgNF) modified electrode. The AgNF was electrodeposited as redox indicator on a gold electrode, which was then functionalized with an electropolymerized layer of PPy, a conducting polymer, to immobilize the PNA probes. The fabrication process was investigated by electrochemical impedance spectroscopy. The biosensor was used to detect miRNA-21, a biomarker abnormally expressed in most cancers. The signal was monitored by the change in current of the AgNF redox reaction before and after hybridization using cyclic voltammetry. Two PNA probe lengths were investigated and the longer probe exhibited a better performance. Nucleotide overhangs on the electrode side affected the signal more than overhangs on the solution side due to the greater insulation of the sensing surface. Under optimal conditions, the electrochemical signal was proportional to miRNA-21 concentrations between 0.20 fM and 1.0 nM, with a very low detection limit of 0.20 fM. The biosensor showed a high specificity which could discriminate between complementary, single-, doubled-base mismatched, and non-complementary targets. Three out of the seven tested plasma samples provided detectable concentrations (63±4, 111±4 and 164±7 fM). The sensor also showed good recoveries (81–119%). The results indicated the possibilities of this biosensor for analysis without RNA extraction and/or amplification, making the sensor potentially useful for both the prognosis and diagnosis of cancer in clinical application.

    更新日期:2017-11-10
  • Nucleic Acid-Based Electrochemical Nanobiosensors
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-04
    Alireza Abi, Zahra Mohammadpour, Xiaolei Zuo, Afsaneh Safavi

    The detection of biomarkers using sensitive and selective analytical devices is critically important for the early stage diagnosis and treatment of diseases. The synergy between the high specificity of nucleic acid recognition units and the great sensitivity of electrochemical signal transductions has already shown promise for the development of efficient biosensing platforms. Yet nucleic-acid based electrochemical biosensors often rely on target amplification strategies (e.g., polymerase chain reactions) to detect analytes at clinically relevant concentration ranges. The complexity and time-consuming nature of these amplification methods impede moving nucleic acid-based electrochemical biosensors from laboratory-based to point-of-care test settings. Fortunately, advancements in nanotechnology have provided growing evidence that the recruitment of nanoscaled materials and structures can enhance the biosensing performance (particularly in terms of sensitivity and response time) to the level suitable for use in point-of-care diagnostic tools. This Review highlights the significant progress in the field of nucleic acid-based electrochemical nanobiosensing with the focus on the works published during the last five years.

    更新日期:2017-11-05
  • Development of an automated wax-printed paper-based lateral flow device for alpha-fetoprotein enzyme-linked immunosorbent assay
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-01
    Pattarachaya Preechakasedkit, Weena Siangproh, Nanthika Khongchareonporn, Nattaya Ngamrojanavanich, Orawon Chailapakul

    In this study, a novel wax-printed paper-based lateral flow device has been developed as an alternative approach for an automated and one-step enzyme-linked immunosorbent assay (ELISA). The design pattern consisted of a non-delayed channel, a wax-delayed channel, a test zone and a control zone. This system was easily fabricated on a nitrocellulose membrane using a wax-printing method and then baked in an oven at 100 °C for 1 min. The four barriers of the wax-delayed channel could delay the flow time for 11 s compared to the flow time of the non-delayed channel. To use the device under optimal conditions, alpha-fetoprotein (AFP) was detected at a limit of detection of 1 ng mL−1 and assessed with the naked eye within 10 min. A colorimetric intensity was also measured using a smart phone and computer software at a linear range of 0.1–100 ng mL−1 with a good correlation. Furthermore, the proposed device was successfully applied to detect AFP in human serum. Therefore, the wax-printing demonstrates a user-friendly, easy and quick method for the fabrication of the device, which could be used as a one-step, portable, disposable, low-cost, simple, instrument-free and point-of-care device for the automated ELISA.

    更新日期:2017-11-02
  • Rh blood phenotyping (D, E, e, C, c) microarrays using multichannel surface plasmon resonance imaging
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-01
    Chinnawut Pipatpanukul, Sasaki Takeya, Akira Baba, Ratthasart Amarit, Armote Somboonkaew, Boonsong Sutapun, Pimpun Kitpoka, Mongkol Kunakorn, Toemsak Srikhirin

    The application of Surface Plasmon Resonance Imaging (SPRi) for the detection of transmembrane antigen of the Rhesus (Rh) blood group system is demonstrated. Clinically significant Rh blood group system antigens, including D, C, E, c, and e, can be simultaneously identified via solid phase immobilization assay, which offers significant time savings and assay simplification. Red blood cells (RBCs) flowed through the micro-channel, where a suitable condition for Rh blood group detection was an RBC dilution of 1:10 with a stop-flow condition. Stop flow showed an improvement in specific binding compared to continuous flow. Rh antigens required a longer incubation time to react with the immobilized antibody than A and B antigens due to the difference in antigen type and their location on the RBC. The interaction between the immobilized antibodies and their specific antigenic counterpart on the RBC showed a significant difference in RBC removal behavior using shear flow, measured from the decay of the SPR signal. The strength of the interaction between the immobilized antibody and RBC antigen was determined from the minimum wall shear stress required to start the decay process in the SPR signal. For a given range of immobilized antibody surface densities, the Rh antigen possesses a stronger interaction than A, B, and AB antigens. Identification of 82 samples of ABO and Rh blood groups using SPRi showed good agreement with the standard micro-column agglutination technique. A wider coverage of antigenic recognition for RBC when using the solid phase immobilization assay was demonstrated for the RBC with the antigenic site located on the transmembrane protein of the clinically significant Rh antigen. Given the level of accuracy and precision, the technique showed potential for the detection of the Rh minor blood group system

    更新日期:2017-11-02
  • A ratiometric electrochemical biosensor for the exosomal microRNAs detection based on bipedal DNA walkers propelled by locked nucleic acid modified toehold mediate strand displacement reaction
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-11-01
    Jing Zhang, Liang-Liang Wang, Mei-Feng Hou, Yao-Kun Xia, Wen-Hui He, An Yan, Yun-Ping Weng, Lu-Peng Zeng, Jing-Hua Chen

    Sensitive and selective detection of microRNAs (miRNAs) in cancer cells derived exosomes have attracted rapidly growing interest owing to their potential in diagnostic and prognostic applications. Here, we design a ratiometric electrochemical biosensor based on bipedal DNA walkers for the attomolar detection of exosomal miR-21. In the presence of miR-21, DNA walkers are activated to walk continuously along DNA tracks, resulting in conformational changes as well as considerable increases of the signal ratio produced by target-respond and target-independent reporters. With the signal cascade amplification of DNA walkers, the biosensor exhibits ultrahigh sensitivity with the limit of detection (LOD) down to 67 aM. Furthermore, owing to the background-correcting function of target-independent reporters termed as reference reporters, the biosensor is robust and stable enough to be applied in the detection of exosomal miR-21 extracted from breast cancer cell lines and serums. In addition, because locked nucleic acid (LNA) modified toehold mediate strand displacement reaction (TMSDR) has extraordinary discriminative ability, the biosensor displays excellent selectivity even against the single-base-mismatched target. It is worth mentioning that our sensor is regenerative and stable for at least 5 cycles without diminution in sensitivity. In brief, the high sensitivity, selectivity and reproducibility, together with cheap, make the proposed biosensor a promising approach for exosomal miRNAs detection, in conjunction with early point-of-care testing (POCT) of cancer.

    更新日期:2017-11-01
  • One-Pot Preparation of Hydrophilic Manganese Oxide Nanoparticles as T1 Nano-Contrast Agent for Molecular Magnetic Resonance Imaging of Renal Carcinoma In Vitro and In Vivo
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-10-27
    Jingjing Li, Chen Wu, Pingfu Hou, Min Zhang, Kai Xu

    Magnetic resonance imaging (MRI) contrast agents have become a necessary part for clinical practice to improve the sensitivity for the diagnosis of small lesions and injuries. Among them, manganese oxide nanoparticle (MnO NPs)-based MRI contrast agent attracts more and more attention because of their better performance in the detection of brain disease and positive enhancement in T1-weighted image. However, the relatively low r1 relaxivity and complex synthetic route hampered their wider applications. In this work, we proposed a one-pot approach to prepare hydrophilic MnO NPs via a polyol-like method with poly (ethylene glycol) (PEG) as both a solvent and surfactant. The obtained PEG-MnO NPs displayed a high T1 relaxivity and a low r2/r1 ratio (12.942 s-1 mM-1 and 4.66) at 3.0 T, which was three times that of the clinical used contrast agent, Magnevist (Gd-DTPA). Additionally, when exposed to the simulated body fluid (SBF), acidic environment or glutathione, PEG-MnO NPs kept stable, favoring their further biological applications. Then, to explore their use for the molecular magnetic resonance imaging of 786-0 renal carcinoma, amino group modified AS1411 aptamer as the targeting molecule was introduced to conjugate with PEG-MnO NPs via covalent coupling reaction. The fabricated nanoprobe, AS1411-PEG-MnO, could clearly visualize 786-0 renal carcinoma cells with MRI in vitro. Furthermore, compared with PEG-MnO NPs, AS1411-PEG-MnO nanoprobe presented a prolonged retention in 786-0 renal carcinoma tumor in vivo. The intravenously injected nanoprobes were eventually excreted from the body through the renal clearance route. These results indicated the potential promising of PEG-MnO NPs as an alternative contrast agent in MRI scanning.

    更新日期:2017-10-28
  • Highly sensitive electrochemical nuclear factor kappa B aptasensor based on target-induced dual-signal ratiometric and polymerase-assisted protein recycling amplification strategy
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-10-26
    Kanfu Peng, Pan Xie, Zhe-Han Yang, Ruo Yuan, Keqin Zhang

    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 0.1 pg mL−1 to 15 ng mL−1 with a low detection limit of 0.03 pg mL−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.

    更新日期:2017-10-27
  • Nanostructured molecularly imprinted polymers for protein chemosensing
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-10-23
    Marcin Dabrowski, Patrycja Lach, Maciej Cieplak, Wlodzimierz Kutner
    更新日期:2017-10-24
  • Detection of Vapor-Phase Organophosphate Threats Using Wearable Conformable Integrated Epidermal and Textile Wireless Biosensor Systems
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-10-20
    Rupesh K. Mishra, Aida Martín, Tatsuo Nakagawa, Abbas Barfidokht, Xialong Lu, Juliane R. Sempionatto, Kay Mengjia Lyu, Aleksandar Karajic, Mustafa M. Musameh, Ilias L. Kyratzis, Joseph Wang
    更新日期:2017-10-20
  • Improved screen-printed carbon electrode for multiplexed label-free amperometric immuniosensor: Addressing its conductivity and reproducibility challenges
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-10-19
    Lihua Zhao, Hongliang Han, Zhanfang Ma

    A new screen-printed carbon electrode (SPCE) with multiple working electrodes and one signal output channel without counter and reference electrodes was designed. The multiple working electrodes can be individually modified for each target of interest. The SPCE contained one signal output channel, making the immuniosensor be realized by common single-channel electrochemical workstation. The counter and reference electrodes were independent of disposable SPCE, reducing costs and eliminating precious metal pollution. Platinum network as counter electrode improved the reproducibility of the SPCE. Moreover, method of generating hydrogel on working electrode was used to enhance the conductivity of SPCE. Based on this, a multiplexed single channel label-free amperometric immuniosensor for four tumor markers, namely, squamous cell carcinoma antigen (SCCA), fragment antigen 21-1 (Cyfra21-1), carbohydrate antigen 125 (CA125), and neuron specific enolase (NSE) was developed, and the corresponding detection limits were 5.5 pg mL−1, 4.8 pg mL−1, 0.0054 U mL−1 and 2.3 pg mL−1, respectively. The sensitivity of this immunosensor was 0.83 µA (lg(ng ml−1))−1 for SCCA, 1.92 µA (lg(ng ml−1))−1 for Cyfra21-1, 4.75 µA (lg(U ml−1))−1 for CA125 and 2.40 µA (lg(ng ml−1))−1 for NSE. Among them, the sensitivities of CA125 and NSE were four-fold higher than those of the previous works.

    更新日期:2017-10-20
  • An Ultrasensitive Electrochemiluminescent Immunosensor Based on Graphene Oxide Coupled Graphite-like Carbon Nitride and Multiwalled Carbon Nanotubes-gold for the Detection of Diclofenac
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-10-19
    Liuyi Hu, Jing Zheng, Kang Zhao, Anping Deng, Jianguo Li

    In this study, a novel competition-type electrochemiluminescent (ECL) immunosensor for detecting diclofenac (DCF) was fabricated with graphene oxide coupled graphite-like carbon nitride (GO-g-C3N4) as signal probe for the first time. The ECL intensity of carboxylated g-C3N4 was significantly enhanced after being combined with graphene oxide (GO) which exhibited excellent charge-transport property. The sensing platform was constructed by multiwalled carbon nanotubes and gold nanoparticles (MWCNTs-AuNPs), which not only provided an effective matrix for immobilizing a large amount of coating antigen but also facilitated the electronic transmission rate to enhance the ECL intensity. Based on the synergistic effect of GO-g-C3N4 and MWCNTs-AuNPs composite, the proposed sensor showed high sensitivity, good stability, and wide linearity for the detection of DCF in the range of 0.005–1000 ng mL−1 with a detection limit of 1.7 pg mL−1. Furthermore, the developed immunoassay has been applied to real samples with satisfactory results. Therefore, this work provided a promising method for the detection of DCF and other small molecular compounds in the future.

    更新日期:2017-10-19
  • Hydrogel micropost-based qPCR for multiplex detection of miRNAs associated with Alzheimer's disease
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-10-19
    Woongsun Choi, Sang Yun Yeom, Junsun Kim, Seungwon Jung, Seungho Jung, Tae Soup Shim, Sang Kyung Kim, Ji Yoon Kang, Soo Hyun Lee, Il-Joo Cho, Jungkyu Choi, Nakwon Choi
    更新日期:2017-10-19
  • Fabrication of Ordered Metallic Glass Nanotube Arrays for Label-free Biosensing with Diffractive Reflectance
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-10-18
    Wei-Ting Chen, May-Show Chen, Jinn P. Chu, Chung-Kwei Lin, Jem-Kun Chen

    In this study, a photoresist template with well-defined contact hole array was fabricated, to which radio frequency magnetron sputtering process was then applied to deposit an alloyed Zr55Cu30Al10Ni5 target, and finally resulted in ordered metallic glass nanotube (MGNT) arrays after removal of the photoresist template. The thickness of the MGNT walls increased from 98 to 126 nm upon increasing the deposition time from 225 to 675 s. The wall thickness of the MGNT arrays also increased while the dimensions of MGNT reduced under the same deposition condition. The MGNT could be filled with biomacromolecules to change the effective refractive index. The air fraction of the medium layer were evaluated through static water contact angle measurements and, thereby, the effective refractive indices the transverse magnetic (TM) and transverse electric (TE) polarized modes were calculated. A standard biotin–streptavidin affinity model was tested using the MGNT arrays and the fundamental response of the system was investigated. Results show that filling the MGNT with streptavidin altered the effective refractive index of the layer, the angle of reflectance and color changes identified by an L⁎a⁎b⁎ color space and color circle on an a⁎b⁎ chromaticity diagram. The limit of detection (LOD) of the MGNT arrays for detection of streptavidin was estimated as 25 nM, with a detection time of 10 min. Thus, the MGNT arrays may be used as a versatile platform for high-sensitive label-free optical biosensing.

    更新日期:2017-10-18
  • An Aptasensor Based on Heparin-Mimicking Hyperbranched Polyester with Anti-Biofouling Interface for Sensitive Thrombin Detection
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-10-18
    Yanlian Niu, Meilin Chu, Ping Xu, Shuangshuang Meng, Qian Zhou, Wenbo Zhao, Bo Zhao, Jian Shen

    In this paper, novel heparin-mimicking hyperbranched polyester nanoparticles (HBPE-SO3 NPs) with abundant of sulfonated acid functional groups were synthesized, and their antithrombogenicities were further evaluated. Further, a label-free electrochemical aptamer biosensor (aptasensor) based on HBPE-SO3 NPs modified electrode was developed for thrombin (TB) detection in whole blood. Meanwhile, the anti-biofouling properties of different modified electrodes were studied by whole blood and platelet adhesion test, hemolysis assay and morphological changes of red blood cells in vitro. Besides, the thrombin-binding aptamer was selected as receptor for the proposed aptasensor, which has excellent binding affinity and selectivity for TB. When binding to TB, the electron transfer taking place at the modified electrode interface was inhibited that can attribute to the stereo-hindrance effect, resulting in the decreased current response. This aptasensor showed excellent electrochemical properties with a wide detection range and a low detection limit of 0.031 pM (S/N=3), and provided high selectivity, long-term stability and good reproducibility. Finally, the sensitively detection of TB in whole blood samples directly was achieved by this aptasensor we proposed, which suggested its great potential for TB detection in the clinic.

    更新日期:2017-10-18
  • Multi-Parametric Rigid and Flexible, Low-Cost, Disposable Sensing Platforms for Biomedical Applications
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-10-18
    Salzitsa Anastasova, Panagiotis Kassanos, Guang – Zhong Yang

    The measurement of Na+, K+ and H+ is essential in medicine and plays an important role in the assessment of tissue ischemia. Microfabrication, inkjet- and screen-printing can be used for solid contact ion selective electrodes (ISE) realization; these, however, can be non-standardized, costly and time consuming processes. We present the realization of ISEs on post-processed electrodes fabricated via standardized printed circuit board (PCB) manufacturing techniques. In vitro results are presented from two rigid platforms (32 ISEs) for liquid sample dip-stick measurements and two flexible platforms (6 and 32 ISEs) for post-surgical intestinal tissue monitoring, each with a common reference electrode (RE). These are combined with optimized tetrapolar bioimpedance sensors for tissue ischemia detection. Both electroless and hard gold PCB finishes are examined. Apart from the electroless rigid platform, the rest demonstrated comparable and superior performance, with the pH sensors demonstrating the greatest deviation; the flexible hard gold platform achieved a sensitivity 4.6 mV/pH and 49.2 mV/pH greater than the electroless flexible and rigid platforms, respectively. The best overall performance was achieved with the hard gold flexible platform with sensitivities as large as 73.4 mV/pH, 56.3 mV/log [Na+], and 57.4 mV/log [K+] vs. custom REs on the same substrate. Simultaneous measurements of target analytes is demonstrated with test solutions and saliva samples. The results exhibit superior performance to other PCB-based pH sensors, demonstration of Na+ and K+ PCB-based sensors with comparable performance to potentiometric sensors fabricated with other techniques, paving the way towards mass-produced, low-cost, disposable, multi-parametric chemical sensing diagnostic platforms.

    更新日期:2017-10-18
  • Sandwich-format ECL immunosensor based on Au star@BSA-Luminol nanocomposites for determination of human chorionic gonadotropin
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-10-18
    Amin Zhang, Weiwei Guo, Hong Ke, Xin Zhang, Huan Zhang, Chusen Huang, Dapeng Yang, Nengqin Jia, Daxiang Cui

    A sandwich-configuration electrochemiluminescence (ECL) immunosensor based on Au star@BSA-Luminol nanocomposites for ultrasensitive determination of human chorionic gonadotropin (HCG) has been developed. In this work, nanostructured Polyaniline hydrogels (Pani) decorated with Pt nanoparticles (Pani/Pt) were utilized to construct the base of this immunosensor, greatly increasing the amount of loaded capture antibodies (Ab1) via linkage reagent glutaraldehyde (GA). The used conducting Pani/Pt nanocomposites possessed the unique features, such as large surface area, high electron transfer speed and favorable electrocatalytic activities of hydrogen peroxide, which offered a prominent platform for this sandwich-sensor and acted as efficient ECL signal amplifier also. Furthermore, we employed horseradish peroxidase (HRP) to block the nonspecific binding sites instead of commonly used bovine serum albumin (BSA), which further amplified the signal of luminol in the present of hydrogen peroxide (H2O2). In addition, Au star@BSA nanocomposites with excellent water solubility, low-toxicity and great biocompatibility were prepared and used to immobilize HCG detection antibodies (Ab2) and luminescent material luminol. Then, the above-synthetized Luminol-Au star@BSA-Ab2 complex was attached to the modified sensor by sandwiched immunoreactions. Under the optimized conditions, the proposed immunosensor exhibited a sensitive detection of HCG in a wide linear range from 0.001–500 mIU mL−1 with a detection limit of 0.0003 mIU/mL (S/N=3). All the results indicated that such a sandwiched HCG immunosensor exhibited favorable ECL analytical performance. This developed method may be potentially used to recognize other clinical protein and display a novel ideal to construct an immunosensor.

    更新日期:2017-10-18
  • Electrochemiluminescent Competitive Immunosensor Based on Polyethyleneimine Capped SiO2 Nanomaterials as Labels to Release Ru(bpy)32+ Fixed in 3D Cu/Ni Oxalate for the Detection of Aflatoxin B1
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-10-18
    Yaoguang Wang, Guanhui Zhao, Xiaojian Li, Li Liu, Wei Cao, Qin Wei

    In this work, a highly-efficient competitive method-based electrochemiluminescence (ECL) immunosensor was proposed based on {[Ru(bpy)3][Cu2xNi2(1-x)(ox)3]}n (Cu/Ni/Ru) as luminophor to efficiently detect aflatoxins B1 (AFB1). Cu/Ni/Ru exhibited excellent ECL behavior. While polyethyleneimine capped silicon dioxide (PEI@SiO2) could decrease the ECL performance of Cu/Ni/Ru due that PEI could destroy the structure of Cu/Ni/Ru via producing the complex between PEI and metal ions (Cu(II)/Ni(II)), inducing the release of Ru(bpy)32+. Since Au nanoparticles can directly combine with antibody and antigen, the Cu/Ni/Ru and PEI@SiO2 was functionalized by Au nanoparticles. The quantitative detection of AFB1 was based on the competitive binding between AFB1-bovine serum albumin labeled Au-PEI@SiO2 (Au-PEI@SiO2-AFB1-BSA) and free AFB1 with antibody-AFB1 which immobilized on Au-Cu/Ni/Ru. The ECL signal increased with augmenting the concentrations of the free AFB1 due to less Au-PEI@SiO2-AFB1-BSA combining with antibodies. Under optimal conditions, the proposed immunosensor exhibited a wide linear range from 0.01 ng mL−1 to 100 ng mL−1 with a detection limit of 0.0039 ng mL−1 (S/N=3). The proposed immunosensor also provides a promising approach for ultrasensitive detection of other mycotoxins.

    更新日期:2017-10-18
  • Perylenetetracarboxylic Acid and Carbon Quantum Dots Assembled Synergistic Electrochemiluminescence Nanomaterial for Ultra-sensitive Carcinoembryonic Antigen Detection
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-10-17
    Lan-lan Xu, Wei Zhang, Lei Shang, Rong-na Ma, Li-ping Jia, Wen-li Jia, Huai-sheng Wang, Li Niu

    It is important to design a nice electrochemiluminescence (ECL) biological nanomaterial for fabricating sensitive ECL immunosensor to detect tumor markers. Most reported ECL nanomaterial was decorated by a number of mono-luminophore. Here, we report a novel ECL nanomaterial assembled by dual luminophores perylenetetracarboxylic acid (PTCA) and carbon quantum dots (CQDs). In the ECL nanomaterial, graphene was chosen as nanocarrier. Significant ECL intensity increases are seen in the ECL nanomaterial, which was interpreted with the proposed synergistic promotion ECL meachanism of PTCA and CQDs. Furthermore, this ECL nanomaterial was used to label secondary antibody and fabricate a sandwiched carcinoembryonic antigen (CEA) immunosensor. The CEA immunosensor exhibits high sensitivity and the linear semilogarithmical range was from 0.001 fg mL-1 to 1 ng mL-1 with low detection limit 0.00026 fg mL-1. And the CEA immunosensor is also suitable for various cancers’ sample detection providing potential specific applications in diagnostics.

    更新日期:2017-10-17
  • Spiral Shape Microfluidic Channel for Selective Isolating of Heterogenic Circulating Tumor Cells
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-10-17
    Bongseop Kwak, Jeonghun Lee, Jaehun Lee, Hyun Soo Kim, Shinwon Kang, Yongkoo Lee

    Detecting heterogenic tumor cells that are traveling in our body through blood stream for the tumor metastasis is one way for cancer prognosis. Due to the heterogeneity of circulating tumor cells (CTCs), further identification of tumor cell types should be accompanied with CTCs isolation from blood cells in peripheral blood sample. Both negative enrichment and recollection of isolated CTCs are required in the downstream analysis, which are time-consuming, labor-intensive, and massive equipment required. To solve these limitations, we have developed a simple and disposable spiral shape microfluidic channel that can separate all CTCs from blood cells, and at the same time, can identify the types of CTCs based on epithelial cell adhesion molecule (EpCAM) expression level. Two different types of tumor cells, MCF-7 and MDA-MB-231, both from the same origin of breast carcinoma cells, were used to demonstrate the functionality of the developed system. The spiral channel system could capture the EpCAM positive and negative CTCs with 96.3% and 81.2% purity, respectively, while both EpCAM positive and negative CTCs were differently positioned along the microfluidic channel. The average selectivity of EpCAM positive and negative CTCs is 6.1:4.8. In addition, the throughput of the system was optimized at a sample flow rate of 150 μl/min. The developed system successfully demonstrated its potential to identify biomarkers, including EpCAM, for detecting the heterogenic CTCs.

    更新日期:2017-10-17
  • A Highly Sensitive VEGF165 Photoelectrochemical Biosensor Fabricated by Assembly of Aptamer Bridged DNA Networks
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-10-17
    Huimei Da, Hongyan Liu, Yingning Zheng, Ruo Yuan, Yaqin Chai

    We developed a novel “signal-off” photoelectrochemical (PEC) aptasensor based on an aptamer bridged DNA network structure for the sensitive detection of vascular endothelial growth factor (VEGF165), using g-C3N4 as photoactive material. The DNA network provides an excellent platform for the immobilization of methylene blue (MB), which can facilitate the electron transport through the DNA helix structure and suppress the recombination of electron-hole pairs generated by g-C3N4. In the presence of the target VEGF165, the DNA network can be destroyed adequately by the recognition between VEGF165 and the aptamer, resulting in the release of MB. Therefore, the originally enhanced electron transfer process could be inhibited, leading to a remarkable decrease of the photocurrent. A good linear relationship between the PEC signal and the logarithm of VEGF165 concentration over the range of 100 fM to 10 nM with a detection limit of 30 fM can be obtained. Our concept can be easily extended to develop aptasensors for the sensitive detection of different targets by triggering the release of the payloads from their corresponding aptamer bridged DNA networks.

    更新日期:2017-10-17
  • Gold atomic cluster mediated electrochemical aptasensor for the detection of Lipopolysaccharide
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-10-16
    Biyas Posha, Sindhu R Nambiar, N. Sandhyarani

    We have constructed an aptamer immobilized gold atomic cluster mediated, ultrasensitive electrochemical biosensor (Apt/AuAC/Au) for LPS detection without any additional signal amplification strategy. The aptamer self-assemble onto the gold atomic clusters makes Apt/AuAC/Au an excellent platform for the LPS detection. Differential pulse voltammetry and EIS were used for the quantitative LPS detection. The Apt/AuAC/Au sensor offers an ultrasensitive and selective detection of LPS down to 7.94×10–21 M level with a wide dynamic range from 0.01 attomolar to 1picomolar. The sensor exhibited excellent selectivity and stability. The real sample analysis was performed by spiking the diluted insulin sample with various concentration of LPS and obtained recovery within 2% error value. The sensor is found to be more sensitive than most of the literature reports. The simple and easy way of construction of this sensor provides an efficient and promising detection of an even trace amount of LPS.

    更新日期:2017-10-17
  • 更新日期:2017-10-17
  • Gold nanopaticle-enhanced multiplexed imaging surface plasmon resonance (iSPR) detection of Fusarium mycotoxins in wheat
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-10-16
    Md Zakir Hossain, Chris M Maragos

    A rapid, sensitive and multiplexed imaging surface plasmon resonance (iSPR) biosensor assay was developed and validated for three Fusarium toxins, deoxynivalenol (DON), zearalenone (ZEA) and T-2 toxin. The iSPR assay was based on a competitive inhibition format with secondary antibodies (Ab2) conjugated to gold nanoparticles (AuNPs) used as a signal amplification tag. Signal was amplified nearly 25-fold for DON, 90-fold for ZEA and 12-fold for T-2 toxin assay using Ab2-AuNPs. Analyses, including steps to regenerate the sensor, took 17.5 min. The antigen coated sensor chip was used for more than 46 cycles without affecting signal intensity (<12%). Matrix matched calibration curves were used to determine Fusarium toxins in wheat. The mean recoveries ranged from 87% to 103% with relative standard deviations of repeatability of less than 5%. The limits of detection were 15 µg/kg for DON, 24 µg/kg for ZEA and 12 µg/kg for T-2 toxin. This provided sufficient sensitivity to monitor contamination of these mycotoxins in wheat in accordance with European Commission (EC) limits. Cut off levels for all three Fusarium toxins were validated using blank wheat and wheat spiked either at the EC regulated levels (100 µg/kg for ZEA and T-2 toxin) or at one third of the EC level (for DON: 400 µg/kg). The assay was successfully applied and further validated with naturally contaminated wheat samples. This is the first reported AuNP enhanced iSPR assay to detect and classify three agriculturally important Fusarium toxins in wheat.

    更新日期:2017-10-17
  • Urinary p-cresol diagnosis using nanocomposite of ZnO/MoS2 and molecular imprinted polymer on optical fiber based lossy mode resonance sensor
    Biosens. Bioelectron. (IF 7.780) Pub Date : 2017-10-16
    Sruthi P. Usha, Banshi D. Gupta

    A lossy mode resonance (LMR) based sensor for urinary p-cresol testing on optical fiber substrate is developed. The sensor probe fabrication includes dip coating of nanocomposite layer of zinc oxide and molybdenum sulphide (ZnO/MoS2) over unclad core of optical fiber as the transducer layer followed by the layer of molecular imprinted polymer (MIP) as the recognition medium. The addition of molybdenum sulphide in the transducer layer increases the absorption of light in the medium which enhances the LMR properties of zinc oxide thereby increasing the conductivity and hence the sensitivity of the sensor. The sensor probe is characterized for p-cresol concentration range from 0 µM (reference sample) to 1000 µM in artificially prepared urine. Optimizations of various probe fabrication parameters are carried to bring out the sensor's optimal performance with a sensitivity of 11.86 nm/µM and 28 nM as the limit of detection (LOD). A two-order improvement in LOD is obtained as compared to the recently reported p-cresol sensor. The proposed sensor possesses a response time of 15 sec which is 8 times better than that reported in the literature utilizing electrochemical method. Its response time is also better than the p-cresol sensor currently available in the market for the medical field. Thus, with a fast response, significant stability and repeatability, the proposed sensor holds practical implementation possibilities in the medical field. Further, the realization of sensor probe over optical fiber substrate adds remote sensing and online monitoring feasibilities.

    更新日期:2017-10-16
Some contents have been Reproduced with permission of the American Chemical Society.
Some contents have been Reproduced by permission of The Royal Society of Chemistry.
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