Article

Printed Highly Ordered Conductive Polymer Nanowires Doped with Biotinylated Polyelectrolytes for Biosensing Applications

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Abstract

The demand for fast and ultratrace biomarkers detection is increasing in bioanalytical chemistry. In this work, highly ordered nanowires array and sensor integration are achieved with nanoscale printing approach. Negatively charged poly(3,4‐ethylenedioxythiophene)–poly(styrenesulfonate) doped with positively charged PEGylated biotin‐derivatized polyelectrolytes results a direct biofunctionalization on the nanowire surface without multiple postmodification steps. It provides homogeneous dispersed biofunctional sites and nonfouling surface on the nanowires. The ordered nanowires array enables the immunosensor to detect biotargets quickly and ultrasensitively. The nanowires impedimetric immunosensor is demonstrated for specific biomarkers detection and achieved a minimum responsive concentration as low as 10 pg mL−1 for protein biomarker and 10 CFU mL−1 for pathogen. A kind of biotin functionalization ink is achieved by doping with PEGylated biotin‐derivatized polyelectrolytes which enables a direct biofunctionalization on the nanowire surface. Then an immunosensor based on highly ordered nanowires array is fabricated by nanoscale printing approach. This sensor can be a general platform for different materials and various bioanalytical applications with ultrahigh sensitivity, trace detection, and fast response.

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... Commercially available conductive polymers such as poly (3, 4-ethylenedioxythiophene)-poly (styrenesulfonate) (PEDOT:PSS) are doped with PEGlated biotin-derivatized polyelectrolytes and printed on the nanowire surface, as shown in Figure 5i. This highly ordered nano-array setup could detect E. coli as low as 10 CFU/mL [167]. Rolling circle amplification (RCA) is a powerful method for DNA amplification, and the authors employed the technique to enhance the detection sensitivity of E. coli O157:H7 in the microfluidic system. ...
... As stated above, detection systems for infectious pathogens have started exploring the potential application of smart materials to develop robust and high throughput technology. A brief summary of the emerging smart materials for microbial pathogens is shown in Table 3. [161] (iv) Wide detection array (v) Relatively large bandwidth terfaces (iv) Tunability of the slowdown factor in given structure Ionic Liquid based systems (i) Both conductor and binder (ii) Good catalytic ability and super sensitivity (iii) High thermal stability (i) Relatively expensive as compared to conventional organic solvents (ii) High cytotoxicity (iii) Mostly limited to electro-analytical system 10 2 CFU/mL [163] 10 3 CFU/mL [164] Responsive Polymer based system (i) Multifunctionality (ii) Structural stability (iii) Facile integration in the detection devices (iv) Tunable detection sensitivity (i) Tedious synthesis process of the designed responsive polymer (ii) Lack of toxicity data profile 10 CFU/mL [167] 10 2 CFU/mL [168] Figure 5. Responsive polymer-based pathogen detection system. (i) (a) Fabrication steps of the immunosensor using conductive polymers such as PEDOT:PSS, and (b) immobilization and detection strategies of E. coli using the nanoarray setup. ...
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... Besides, the increased surface area by the nanoscale electrodes improves the sensitivity as well as signal to noise ratio of the sensor (Liu et al., 2017). The molecular design of the bio-ink was introduced in detail in our previous work (see also the Supplementary Materials S1) (Xue et al., 2019). Poly(3,4-ethylenedioxythiophene) polystyrene sulfonate J o u r n a l P r e -p r o o f (PEDOT:PSS) is negatively charged and polyelectrolytes (PLL-g-OEG4-Biotin) is positively charged, which leads to the direct formation of bio functionalized nanowires without multiple post modification steps ( Figure s1). ...
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... To evaluate the thermodynamic favorability for the formation of various supramolecular associates in the chemical systems under study, we performed quantum chemical calculations using density functional theory (DFT) (for details, see the Computational Details section and Supporting Information, Tables S4 and S5). The results of our quantum chemical calculations reveal that (i) self-assembly of PDADMAC and heparin to dimeric associates heparin/PDADMAC is thermodynamically profitable (by 43 6 ] 2+ is dramatic, and removal of small imaginary frequencies does not change the thermodynamics estimation qualitatively. ...
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Self-assembled nanowire (NW) crystals can be grown into nearly defect-free nanomechanical resonators with exceptional properties, including small motional mass, high resonant frequency, and low dissipation. Furthermore, by virtue of slight asymmetries in geometry, a NW's flexural modes are split into doublets oscillating along orthogonal axes. These characteristics make bottom-up grown NWs extremely sensitive vectorial force sensors. Here, taking advantage of its adaptability as a scanning probe, we use a single NW to image a sample surface. By monitoring the frequency shift and direction of oscillation of both modes as we scan above the surface, we construct a map of all spatial tip-sample force derivatives in the plane. Finally, we use the NW to image electric force fields distinguishing between forces arising from the NW charge and polarizability. This universally applicable technique enables a form of atomic force microscopy particularly suited to mapping the size and direction of weak tip-sample forces.
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Development of high-performance p-type semiconductor based gas sensors exhibiting fast-response/recovery times with ultra-high response are of major importance for gas sensing applications. Recent reports demonstrated the excellent properties of p-type semiconducting oxide for various practical applications, especially for selective oxidation of volatile organic compounds (VOCs). In this work, sensors based on CuO nanowire (NW) networks have been successfully fabricated via a simple thermal oxidation process on pre-patterned Au/Cr pads. Our investigation demonstrates high impact of the process temperature on aspect ratio and density of copper oxide NWs. An optimal temperature for growth of thin and densely packed NWs was found to be at 425 °C. The fabricated sensors demonstrated ultra-high gas response by a factor of 313 to ethanol vapour (100 ppm) at an operating temperature of 250 °C. High stability and repeatability of these sensors indicate the efficiency of p-type oxide based gas sensors for selective detection of VOCs. A high-performance nanodevice was fabricated in a FIB-SEM system using a single CuO NW, demonstrating an ethanol response of 202 and rapid response and recovery of ~198 ms at room temperature. The involved gas sensing mechanism of CuO NW networks has been described. We consider that the presented results will be of a great interest for the development of higher-performance p-type semiconductor based sensors and bottom-up nanotechnologies.
Article
In this study, different morphological ZnO nanostructures, those of sharp nanowires (NWs), rod NWs, and hexahedral-puncheon nanostructures, were grown in microfluidic channels on the same glass substrate. Characterizations of correspondent biomolecule binding properties were simulated and demonstrated. The surface was modified using 3-ammineopropyl-triethoxysilane (3-APTES) and biotin-N-hydroxysuccinimide ester (NHS-biotin). Different concentrations (4.17pM to 41.7nM) of dye-conjugated streptavidin were simultaneously infused through the second microfluidic channels, which lie 90° from the first microfluidic channels. The florescent intensity at the crossover areas showed good agreement with simulations, with sharp ZnO NWs exhibiting the largest dynamic range and the highest fluorescent intensity. We further characterize correspondent protein detection using sharp ZnO NWs. The surfaces of these ZnO NWs were modified with mouse immunoglobulin G (IgG), infused through the second microfluidic channels with dye-conjugated (Alexa 546) anti-mouse IgG in different concentrations. Concentrations ranging from 417fM to 41.7nM can be resolved using sharp ZnO NWs. Finally, multiple protein detection was demonstrated using a five-by-eight microfluidic channel array. Fluorescence images present clear multiple detections at the crossover areas when using the sharp ZnO NWs for simultaneous dye-conjugated anti-mouse IgG and dye-conjugated anti-rabbit IgG (Alexa 647) detection. Copyright © 2015 Elsevier B.V. All rights reserved.
Article
Nanoscale circuits are fabricated by assembling different conducting materials (e.g., metal nanoparticles, metal nano-wires, graphene, carbon nanotubes, and conducting polymers) on inkjet-printing patterned substrates. This nonlitho-graphy strategy opens a new avenue for integrating conducting building blocks into nanoscale devices in a cost-efficient manner. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Article
A new surface functionalization scheme for nano-Bio field effect transistors (FETs) using biocompatible polyelectrolyte thin films (PET) is developed. PET assemblies on Si nanowires (Si-NWs) are driven by electrostatic interactions between the positively charged polymer backbone and negatively charged Si/SiO2 surface. Such assemblies can be directly coated from PET aqueous solutions and result in a uniform nanoscale thin film, which is more stable compared to the conventional amine silanization. Short oligo-ethylene glycol chains are grafted on the PETs to prevent nonspecific protein binding. Moreover, the reactive groups of the polymer chains can be further functionalized to other chemical groups in specific stoichiometry for biomolecules detection. Therefore, it opens a new strategy to precisely control the functional group densities on various biosensor surfaces at the molecular level. In addition, such assemblies of the polymers together with the bound analytes can be removed with the pH stimulation resulting in regeneration of a bare sensor surface without compromising the integrity and performance of the Si-NWs. Thus, it is believed that the developed PET coating and sensing systems on Si-NW FETs represent a versatile, promising approach for regenerative biosensors which can be applied to other biosensors and will benefit real device applications, enhancing sensor lifetime, reliability, and repeatability.
Article
We report here the use of nanomolding in capillaries (NAMIC) coupled with dithiocarbamate (DTC) chemistry to fabricate sub-50 nm quasi-1D arrays of 3.5 nm core gold nanoparticles (Au NPs) over large areas. Owing to chemical immobilization via the DTC bond, the patterned NP systems are stable in water and organic solvents, thus allowing the surface modification of the patterned Au NP arrays through thiol chemistry and further orthogonal binding of proteins. The electrical properties of these patterned Au NP wires have also been studied. Our results show that NAMIC combined with surface chemistry is a simple but powerful tool to create metal NP arrays that can potentially be applied to fabricate nanoelectronic or biosensing devices.
Article
Nanowires are important potential candidates for the realization of the next generation of sensors. They offer many advantages such as high surface-to-volume ratios, Debye lengths comparable to the target molecule, minimum power consumption, and they can be relatively easily incorporated into microelectronic devices. Accordingly, there has been an intensified search for novel nanowire materials and corresponding platforms for realizing single-molecule detection with superior sensing performance. In this work, progress made towards the use of nanowires for achieving better sensing performance is critically reviewed. In particular, various nanowires types (metallic, semiconducting, and insulating) and their employment either as a sensor material or as a template material are discussed. Major obstacles and future steps towards the ultimate nanosensors based on nanowires are addressed.
Article
We have developed a simple assay method for the evaluation of estrogen receptor (ER) binding capacity of chemicals without the use of radio- or fluorescence-labeled compounds. We used the solution competition assay by the BIACORE biosensor, a surface plasmon resonance biosensor, with estradiol as a ligand, human recombinant ER(alpha) (hrER(alpha)) as a high molecular weight (hmw) interactant and test chemicals as analytes. For the ligand, aminated estradiol with a spacer molecule (E2-17PeNH) was synthesized and immobilized on a carboxymethyl dextran-coated sensor chip by the amine coupling method. The injection of the hmw interactant hrER(alpha) to the biosensor raised the sensorgram, indicating its binding to the ligand E2-17PeNH. The binding of test chemicals to hrERalpha was determined as a reduction in the hrER(alpha) binding to E2-17PeNH. The dissociation constant for the binding to hrER(alpha) was calculated for estrone (4.29 x 10(-9)M), estradiol (4.04 x 10(-10)M), estriol (8.35 x 10(-10)M), tamoxifen (2.16 x 10(-8)M), diethylstilbestrol (1.46 x 10(-10)M), bisphenol A (1.35 x 10(-6)M) and 4-nonylphenol (7.49 x 10(-6)M), by plotting the data according to an equation based on mass action law. This method can also be used as a high throughput screening method.
Article
Avidin-poly(ethylene glycol) (PEG) conjugates were obtained by derivatization of about 10% of the protein amino groups (four amino groups per protein molecule) with linear 5 kDa PEG or branched 10 or 20 kDa PEGs. Circular dichroism analysis showed that the polymer conjugation neither altered the protein structure nor the environment of the aromatic amino acids which are present at the level of the biotin binding site. Spectroscopic studies were carried out to evaluate the biotin recognition activity of the conjugates either in terms of number of biotin binding sites or avidin/biotin affinity. Avidin-PEG 5 kDa and avidin-PEG 10 kDa displayed over 90% of the native protein biological activity while a reduction in the recognition of biotinylated antibodies of about 25% was found with PEG 20 kDa. In vivo studies demonstrated that the protein immunogenicity was in the order: wild type avidin>avidin-PEG 5 kDa>avidin-PEG 10 kDa>avidin-PEG 20 kDa. By intravenous injection into mice bearing a solid tumor, all conjugates displayed prolonged permanence in the circulation with respect to the native protein. The area under the curve values of avidin-PEG 5 kDa, avidin-PEG 10 kDa and avidin-PEG 20 kDa were about 3-, 7- and 30-times higher than the wild type avidin with reduced accumulation in kidneys and liver. Interestingly, all conjugates accumulated significantly in the tumor mass. In particular, in the case of avidin-PEG 20 kDa, 8% of the injected dose (ID)/g of tissue accumulated in the tumor after 5 h from the administration and over 6% of the ID/g was maintained throughout 72 h.
Article
In order to establish ELISA (enzyme-linked immunosorbent assay) method to detect Total E. coli in water environment, E. coli multi-characters antigens in water environment were prepared according to the characters of kinds of E. coli serotypes, including antigen of whole cell, antigen of disrupted whole cell, somatic antigen, flagellar antigen and fimbrial antigen. Total E. coli polyclonal antibodies were obtained from the New Zealand rabbits immunized with these five antigens, respectively. Antibodies generated in this research are with high titers and good purity, can conjugate with antigens, specifically, stably and strongly. Indirect ELISA shows the titers of antibody of whole cell and antibody of disrupted whole cell are both over 1x10(5). The cross-reactivity of the antibody is from 12 to 30% which indicate the specificity of the antibody against Total E. coli. Based on these antibodies, we established indirect ELISA method to detect Total E. coli in water environment. The matrix effects were studied and the results show that there is no significant influence by all the factors. The ELISA result shows that the detection limitation could be 10(4) CFU (colony forming units) L(-1). The indirect ELISA method developed in this study is well suited for Total E. coli analysis in real water samples as a rapid screen method.
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