Dendrimer-Au Nanoparticle Network Covered Alumina Membrane for Ion Rectification and Enhanced Bioanalysis.

Ion transport in an artificial asymmetric nanoporous membrane, which is similar to biological ion channels, can be used for biosensing. Here, a dendrimer-Au nanoparticle network (DAN) is in situ assembled on a nanoporous anodic aluminum oxide (AAO) surface, forming a DAN/AAO hybrid membrane. Benefiting from the high surface area and anion selectivity of DAN, the prepared DAN/AAO hybrid presents selective ion transport. Under a bias potential, a diode-like current-potential (I-V) response is observed. The obtained ionic current rectification (ICR) property can be tuned by the ion valence and pH value of the electrolyte. The rectified ionic current endows the as-prepared DAN/AAO hybrid with the ability of enhanced bioanalysis. Sensitive capture and detection of circulating tumor cells (CTCs) with a detection limit of 80 cells mL-1 as well as excellent reusability can be achieved.

Water as a Universal Infrared Probe for Bioanalysis in Aqueous Solution by Attenuated Total Reflection-Surface Enhanced Infrared Absorption Spectroscopy.

Monitoring the properties and reactions of biomolecules at their interface has attracted ever-growing interest. Here, we propose an approach of infrared analysis technique that utilizes water molecule as a universal probe for in situ and label free monitoring of interfacial bioevents in aqueous solution with high sensitivity. The strong infrared (IR) signal of O-H stretching vibrations from the repelled water is used to sensitively reveal the kinetics of interfacial bioevents at molecular level based on the steric displacement of water using an attenuated total reflection-surface enhanced infrared absorption spectroscopy. Using interfacial immuno-recognition and DNA hybridization as demonstrations, water IR probe offers 26 and 34 times higher sensitivity and even 200 and 86 times lower detection limit for immunosensing and DNA sensing, respectively, as compared to the traditional IR molecular fingerprints.

Enhanced Peroxidase-Like Performance of Gold Nanoparticles by Hot Electrons.

Enzyme mimics have been widely used as alternatives to natural enzymes. However, the catalytic performances of enzyme mimics are often decreased due to different spatial structures or absence of functional groups compared to natural enzymes. Here, we report a highly efficient enzyme-like catalytic performance of gold nanoparticles (AuNPs) by visible-light stimulation. The enzyme-like reaction is evaluated by the catalytic reaction of AuNPs oxidizing a typical chromogenic substrate 3,3',5,5'-tetramethylbenzydine (TMB) with hydrogen peroxide as an oxidant. From investigations of the wavelength-dependent reaction rate, radical capture, hole-donor addition, and dark-field scattering spectroscopy experiments, it is revealed that the strong plasmonic absorption of AuNPs facilitates generation of hot electrons, which are transfered from AuNPs to the adsorbed reactant molecule, greatly promoting the catalytic performance of the enzyme-like catalytic reaction. The present work provides a simple method for improving the performance of enzyme mimics, which is expected to find further application in the field of plasmon-enhanced biocatalysis and biosensors.

Fluorescent Sulfur-Tagged Europium(III) Coordination Polymers for Monitoring Reactive Oxygen Species.

Oxidative stress caused by reactive oxygen species (ROS) is harmful to biological systems and implicated in various diseases. A variety of selective fluorescent probes have been developed for detecting ROS to uncover their biological functions. Generally, the preparation of the fluorescent probes usually undergoes multiple synthetic steps, and the successful fluorescent sensing usually relies on trial-and-error tests. Herein we present a simple way to prepare fluorescent ROS probes that can be used both in biological and environmental systems. The fluorescent europium(III) coordination polymers (CPs) are prepared by simply mixing the precursors [2,2'-thiodiacetic acid and Eu(NO3)3·6H2O] in ethanol. Interestingly, with the increase of reaction temperature, the product undergoes a morphological transformation from microcrystal to nanoparticle while the structure and fluorescent properties retain. The fluorescence of the sulfur-tagged europium(III) CPs can be selectively quenched by ROS, and thus, sensitive and selective monitoring of ROS in aerosols by the microcrystals and in live cells by the nanoparticles has been achieved. The results reveal that the sulfur-tagged europium(III) CPs provide a novel sensor for imaging ROS in biological and environmental systems.

Fenton reaction-mediated fluorescence quenching of N-acetyl-L-cysteine-protected gold nanoclusters: analytical applications of hydrogen peroxide, glucose, and catalase detection.

Given the importance of hydrogen peroxide (H2O2) in many biological processes and its wide application in various industries, the demand for sensitive, accurate, and economical H2O2 sensors is high. In this study, we used Fenton reaction-stimulated fluorescence quenching of N-acetyl-L-cysteine-protected gold nanoclusters (NAC-AuNCs) as a reporter system for the determination of H2O2. After the experimental conditions were optimized, the sensing platform enabled the analysis of H2O2 with a limit of detection (LOD) as low as 0.027 µM. As the glucose oxidase cascade leads to the generation of H2O2 and catalase catalyzes the decomposition of H2O2, these two biocatalytic procedures can be probed by the Fenton reaction-mediated quenching of NAC-AuNCs. The LOD for glucose was found to be 0.18 µM, and the linear range was 0.39-27.22 µM. The LOD for catalase was 0.002 U mL(-1), and the linear range was 0.01-0.3 U mL(-1). Moreover, the proposed sensing methods were successfully applied for human serum glucose detection and the non-invasive determination of catalase activity in human saliva, demonstrating their great potential for practical applications.

Donnan potential caused by polyelectrolyte monolayers.

The Donnan potential is successfully isolated from ion pair potential on a ferrocene-labeled polyelectrolyte (DNA) monolayer. The isolated Donnan potential shifts negatively upon the increase in NaClO4 concentration with a slope of -58.8 mV/decade. With the salt concentration grown up to 1 M, the stretched DNA chains in low salt concentration are found to experience a gradual conformation relaxing process. At salt concentrations higher than 2 M, Donnan breakdown occurs where only the ion pair effect modulates the apparent potential. The apparent formal potential also shows strong dependence on solution pH, which reveals that the charge density in the polyelectrolyte monolayer plays an important role in the establishment of Donnan equilibrium.

Label-free strategy for in-situ analysis of protein binding interaction based on attenuated total reflection surface enhanced infrared absorption spectroscopy (ATR-SEIRAS).

A versatile ATR-SEIRAS methodology is described herein for highly sensitive analysis of immunoglobulin (IgG) recognition. This strategy allows in situ tracking of specific protein binding at the liquid-solid interface. Most importantly, interferential signal from environmental molecules (e.g., water, nonspecific binding molecules, and bulk molecules) can be eliminated to negligible levels by using the ATR analysis mode, and the sensitive IR structural information of target proteins is obtained simultaneously. A simplified numerical model has been established to quantitatively describe the kinetics and thermodynamics of protein recognition processes at surfaces. Compared with conventional label-free methods for protein binding study, experimental results obtained from IR spectroscopic information are more reliable. The presented ATR-SEIRAS method is powerful in studying surface limited protein binding reactions.

RETRACTED: Liposomal valinomycin mediated cellular K+ leak promoting apoptosis of liver cancer cells.

This article has been retracted: please see Elsevier Policy on Article Withdrawal (https://www.elsevier.com/about/our-business/policies/article-withdrawal). This article has been retracted at the request of the corresponding author. It has been found that Fig 2B contains manipulated components, and Fig 5A partially overlaps with Fig 6 of a published paper authored by Mirza Muhammad Faran Ashraf Baig, et, al., The effective transfection of a low dose of negatively charged drug-loaded DNA-nanocarriers into cancer cells via scavenger receptors, J. Pharm. Anal. 11 (2021) 174-182, https://doi.org/10.1016/j.jpha.2020.10.003. The corresponding author indicated that they cannot guarantee the integrity of the images in the manuscript, as well as the conclusions of the paper. As a result, the Editor-in-Chief has decided to retract the paper. The corresponding author deeply regrets the circumstances and apologizes to the scientific community for not having detected this prior to publication.

Living-DNA Nanogel Appendant Enables In Situ Modulation and Quantification of Regulation Effects on Membrane Proteins.

Screening appendants on membrane proteins to understand their varied regulation effects is desirable for finding the potential candidates of the membrane-protein-targeted drugs. However, most artificial appendants can hardly support in situ condition screening because they cannot evolve in situ, neither can they send out signals to reflect the modulation. Here, we designed living-DNA appendants to enable such screening. First, the living-cell rolling-circle amplification (LCRCA) strategy was developed to elongate the DNA appendants for self-tangled physical nanogels. The nanogels unify both the functions of membrane-protein modulation and quantification: their sizes increase with the increased time length of LCRCA, which change the regulation effect on the membrane proteins; their large number of repeating short sequences allow quantification of their sizes in the presence of the complementary fluorophore-tagged short DNA. Then, the performance of the living-DNA appendants was examined taking α6ß4 integrins as the target, where effective regulation over the distribution of actin filaments, cell viability, and chances of anoikis are all validated. The screening also clearly elucidates the interesting nonlinear relationships between the regulations and the effects. We hope this screening strategy based on living-DNA appendants can stand for a prototype for deeper understanding of natural behaviors of membrane proteins and help in the accurate designing of the membrane-protein-targeted drugs.

Protein-Supported RuO2 Nanoparticles with Improved Catalytic Activity, In Vitro Salt Resistance, and Biocompatibility: Colorimetric and Electrochemical Biosensing of Cellular H2O2.

Protein-supported nanoparticles have a great significance in scientific and nanotechnology research because of their "green" process, low cost-in-use, good biocompatibility, and some interesting properties. Ruthenium oxide nanoparticles (RuO2NPs) have been considered to be an important member in nanotechnology research. However, the biosynthetic approach of RuO2NPs is relatively few compared to those of other nanoparticles. To address this challenge, this work presented a new way for RuO2NP synthesis (BSA-RuO2NPs) supported by bovine serum albumin (BSA). BSA-RuO2NPs are confirmed to exert peroxidase-like activity, electrocatalytic activity, in vitro salt resistance (2 M NaCl), and biocompatibility. Results indicate that BSA-RuO2NPs have higher affinity binding for 3,3',5,5'-tetramethylbenzidine or H2O2 than bare RuO2NPs. Moreover, BSA turns out to be a crucial factor in promoting the stability of RuO2NPs. Taking the advantages of these improved properties, we established colorimetric (linear range from 2 to 800 µM, a limit of detection of 1.8 µM) and electrochemical (linear range from 0.4 to 3850 µM, a limit of detection of 0.18 µM) biosensors for monitoring in situ H2O2 secretion from living MCF-7 cells. Herein, this work offers a new biosynthesis strategy to obtain BSA-RuO2NPs and sheds light on the sensitive biosensors to monitor the H2O2 secreted from living cells for promising applications in the fields of nanotechnology, biology, biosensors, and medicine.

Rapidly Visualizing the Membrane Affinity of Gene Vectors Using Polydiacetylene-Based Allochroic Vesicles.

The high-throughput screening of chemically active substances has aroused widespread interest in recent years. The screening of drug carriers is usually ignored, although they interact directly with physiological barriers and target cells, and they determine the fate and efficacy of drugs in vivo. In this work, a series of polydiacetylene (PDA) vesicles (ca. 550 nm) that simulate the cell membrane are constructed to detect the membrane affinity of gene vectors. The surface potentials of vesicles are adjusted by changing the phospholipid composition using different charged compounds. All vesicles show the rapid color changes upon the addition of gene vectors by the naked eye within <5 min. The optimized 1,2-dimyristoyl- sn-glycero-3-phosphocholine (DMPC)-PDA vesicles display the most sensitive discoloration response to the commercially available gene vectors, including Lipofectamine 2000, Entranster-H4000, and polyethylenimine. The logarithm of transfection efficiency for green fluorescent protein plasmid (pGFP) mediated by these three vectors in L02 and HepG2 cells demonstrate an excellent linear correlation with the logarithm of membrane affinity (log Kb) of the gene vectors detected by DMPC-PDA vesicles. This rapid visualization method not only allows the in vitro membrane affinity prediction of gene vectors that greatly contributes to the gene transfection efficiency, but also offers a universal strategy for the potential high-throughput screening of various carrier materials featuring high cell affinity.

Hemoglobin on phosphonic acid terminated self-assembled monolayers at a gold electrode: immobilization, direct electrochemistry, and electrocatalysis.

Phosphonic acid (--PO(3)H(2)) terminated self-assembled monolayers (SAMs) on a gold surface were used as a functional interface to immobilize hemoglobin (Hb). In situ surface-enhanced infrared absorption spectroscopy (SEIRAS) measurements show that Hb immobilization is a sluggish process due to formation of multilayer Hb structures on the PO(3)H(2)-terminated SAMs, as revealed by ellipsometry, atomic force microscopy (AFM), and cyclic voltammetry (CV). In the multilayered Hb film, the innermost Hb molecules can directly exchange electrons with the electrode, whereas Hb beyond this layer communicates electronically with the electrode via protein-protein electron exchange. In addition, electrochemical measurements indicate that immobilization of Hb on the PO(3)H(2)-terminated SAMs is not driven by the electrostatic interaction, but likely by hydrogen-bonding interaction. The immobilized Hb molecules show excellent bioelectrocatalytic activity towards hydrogen peroxide, that is, the PO(3)H(2)-terminated SAMs are promising for construction of third-generation biosensors.

Simultaneous voltammetric determination of norepinephrine, ascorbic acid and uric acid on polycalconcarboxylic acid modified glassy carbon electrode.

A novel polycalconcarboxylic acid (CCA) modified glassy carbon electrode (GCE) was fabricated by electropolymerization and then successfully used to simultaneously determine ascorbic acid (AA), norepinephrine (NE) and uric acid (UA). The characterization of electrochemically synthesized Poly-CCA film was investigated by atomic force microscopy (AFM), electrochemical impedance spectroscopy (EIS) and voltammetric methods. It was found that the electrochemical behavior of the polymer-modified electrode depended on film thickness, i.e., the electropylmyerization time. Based on the electrochemical data, the charge transfer coefficient (alpha) and the surface coverage (Gamma) were calculated. This poly-CCA modified GCE could reduce the overpotential of ascorbic acid (AA), norepinephrine (NE) and uric acid (UA) oxidation in phosphate buffer solution (pH 6.0), while it increases the peak current significantly. The current peak separations of AA/NE, NE/UA and AA/UA on this modified electrode are 91mV, 256mV and 390mV in CV at 100mVs(-1), respectively. Therefore, the voltammetric responses of these three compounds can be well resolved on the polymer-modified electrode, and simultaneously determination of these three compounds can be achieved. In addition, this modified electrode can be successfully applied to determine AA and NE in injection and UA in urine samples without interferences.

In situ synthesis and characterization of multi-walled carbon nanotube/Prussian blue nanocomposite materials and application.

An effective and facile in situ electroless deposition approach for the fabrication of multi-walled carbon nanotube-supported Prussian blue nanoparticle (MWNTs/PB NP) composite nanomaterials is demonstrated in this article. The coverage of PB NPs on MWNTs is tunable by varying the experimental parameters, such as the initial molar concentration of FeCl3 + K3[Fe(CN)6], the relative concentration of FeCl3 + K3[Fe(CN)6] to MWNTs, and the temperature and duration of the heat treatment. This method involves a simple mixing process followed by a mild heating process and does not need the exhaustive surface oxidation process of MWNTs. TEM, FTIR, UV, and XRD are all used to characterize the MWNTs/PB composite materials. In addition, the electrochemical behavior of PB and catalysis of the reduction of H2O2, are investigated. The novel method is expected to be applicable for preparation of other coordination polymer/MWNTs composites and finds use in applications for electronic nanodevices.

Off-line form of the Michaelis-Menten equation for studying the reaction kinetics in a polymer microchip integrated with enzyme microreactor.

We firstly transformed the traditional Michaelis-Menten equation into an off-line form which can be used for evaluating the Michaelis-Menten constant after the enzymatic reaction. For experimental estimation of the kinetics of enzymatic reactions, we have developed a facile and effective method by integrating an enzyme microreactor into direct-printing polymer microchips. Strong nonspecific adsorption of proteins was utilized to effectively immobilize enzymes onto the microchannel wall, forming the integrated on-column enzyme microreactor in a microchip. The properties of the integrated enzyme microreactor were evaluated by using the enzymatic reaction of glucose oxidase (GOx) with its substrate glucose as a model system. The reaction product, hydrogen peroxide, was electrochemically (EC) analyzed using a Pt microelectrode. The data for enzyme kinetics using our off-line form of the Michaelis-Menten equation was obtained (K(m) = 2.64 mM), which is much smaller than that reported in solution (K(m) = 6.0 mM). Due to the hydrophobic property and the native mesoscopic structure of the poly(ethylene terephthalate) film, the immobilized enzyme in the microreactor shows good stability and bioactivity under the flowing conditions.

Versatile microfluidic droplets array for bioanalysis.

We propose a novel method to obtain versatile droplets arrays on a regional hydrophilic chip that is fabricated by PDMS soft lithography and regional plasma treatment. It enables rapid liquid dispensation and droplets array formation just making the chip surface in contact with solution. By combining this chip with a special Christmas Tree structure, the droplets array with concentrations in gradient is generated. It possesses the greatly improved performance of convenience and versatility in bioscreening and biosensing. For example, high throughput condition screening of toxic tests of CdSe quantum dots on HL-60 cells are conducted and cell death rates are successfully counted quickly and efficiently. Furthermore, a rapid biosensing approach for cancer biomarkers carcinoma embryonic antigen (CEA) is developed via magnetic beads (MBs)-based sandwich immunoassay methods.

Electrochemical detector for microchip electrophoresis of poly(dimethylsiloxane) with a three-dimensional adjustor.

This paper presents an electrochemical detector for poly(dimethylsiloxane) (PDMS) microchip CE with a three-dimensional adjustor which makes it possible to accurately align a separate working electrode that can be easily fabricated in laboratory to the uncertain PDMS microchannel outlet. The substantial influence of the electrode-PDMS microchannel distance was investigated. The optimal electrode-outlet distance was found to be 10 microm for the PDMS microchannel with the width of 50 microm due to its relatively slow electroosmotic flow. Adrenaline and catechol were well separated, with a linear response range from 20 microM to 1 mM, and a detection limit of 2 microM for catechol, using carbon disk electrode (diameter of 300 microm). Furthermore, arginine and histidine can be well separated and detected directly in the PDMS microchannel using a Cu disk electrode (diameter of 150 microm).

Remote control of reversible localized protein adsorption in microfluidic devices.

We present a facilely prepared graphene oxide (GO)/ poly(dimethylsiloxane) (PDMS) composite by dispersing nanosized GO in PDMS. On the basis of the combination of photothermal effects of GO and grafted thermoresponsive polymer, poly(N-isopropylacrylamide) (PNIPAAm), an optical-driving approach for remote control of localized wettability is realized. And this method has been successfully applied in the spatially controlled reversible protein adsorption in microfluidic devices.

Sensitive determination of reactive oxygen species in cigarette smoke using microchip electrophoresis-localized surface plasmon resonance enhanced fluorescence detection.

A sensitive approach to the determination of reactive oxygen species (ROS) in puffs of cigarette smoke (CS) has been developed. The experimental system consists of a microfluidic chip electrophoresis and a laser induced fluorescence (LIF) device enhanced by localized surface plasmon resonance. Core-shell Ag@SiO2 nanoparticles were prepared and then immobilized on the surface of the microchannel to increase the fluorescence intensity based on localized surface plasmon resonance-enhanced fluorescence (LSPREF) effect. The ROS in puffs of CS were trapped via the oxidation of 2',7'-dichlorodihydrofluorescein (DCHF) that had been loaded on polyacrylonitrile (PAN) nanofibers in a micro-column. Determination of ROS was based on the amount of 2',7'-dichlorofluorescein (DCF), which is the sole product from DCHF oxidation. With the optimization of the trapping efficiency, we detected about 8.0 pmol of ROS per puff in the mainstream CS. This microchip electrophoresis-SPREF system enables sensitive quantitation of ROS in CS with low consumption of reagent, material, and analysis time.

On-chip selective capture of cancer cells and ultrasensitive fluorescence detection of survivin mRNA in a single living cell.

The rapid recognition of cancer cells and detection of tumor biomarker survivin mRNA plays a critical role in the early diagnosis of many cancers. Based on the integration of specific cancer cell capture and intracellular survivin mRNA detection, this work presents a novel and sensitive on-chip approach for the bioanalysis of survivin mRNA in a single living cell. The microchannel surface was firstly modified with a prostate stem cell antigen (PSCA) monoclonal antibody as the recognition element for prostate cancer cells (PC-3). As a result of the antigen-antibody specific affinity interactions, PC-3 cells could be selectively captured on the microchannel surface. After cell capture, nano-sized graphene oxide-poly(ethylene glycol) bis(amine) (NGO-PEG) was employed as a quencher and carrier of a signal tag, fluorescein isothiocyanate (FITC)-labeled antisense oligonucleotide (F-S1), which is complementary to part of survivin mRNA (target survivin mRNA), to transfect into the captured PC-3 cells. Upon the selective binding of S1 to intracellular survivin mRNA, F-S1 will be released from the NGO-PEG, inducing the fluorescence recovery of FITC. This antibody-based microfluidic device enables simple and inexpensive monitoring of the amount of survivin mRNA in single captured cell without the need for sample pretreatment. The survivin mRNA content in each PC-3 cell was estimated to be (4.8 ± 1.8) × 10(6) copies. This strategy opens a different perspective for ultrasensitive survivin mRNA detection, which may facilitate the early screening for malignancy.