Chusen Huang

Highly sensitive electrochemical impedance spectroscopy immunosensor for the detection of AFB1 in olive oil

Aflatoxin produced by Aspergillus flavus and Aspergillus parasiticus are commonly found in olive and its derivatives. Aflatoxin B1 (AFB1) is a predominant toxin detected abundantly and has been implicated in the etiology of human hepatocellular carcinoma. This study proposes a sensitive and convenient electrochemical impedance spectroscopy (EIS) method for determining AFB1 by MWCNTs/RTIL composite films-based immunosensor. The calibration curve for AFB1 was linear in the range of 0.1-10ngmL(-1) with the limit of detection (LOD) 0.03ngmL(-1). The presence of MWCNTs warrant fast electron transfer, and the ionic liquid provides a benign microenvironment for antibody. The experimental parameters, such as pH and incubating time, have been investigated and optimized. Furthermore, the detection of AFB1 is presented to test this method after extracted from olive oils. It can be anticipated that this method would be used for the detection of AFB1 in various agriculture products and vegetable oils.

Sensitive ECL immunosensor for detection of retinol-binding protein based on double-assisted signal amplification strategy of multiwalled carbon nanotubes and Ru(bpy)32+ doped mesoporous silica nanospheres

A novel electrochemiluminescence (ECL) strategy based on the sandwich-type immunosensor for sensitive detection of retinol-binding protein (RBP) was developed. The primary antibody anti-RBP was immobilized onto multiwalled carbon nanotubes (MWCNTs), which have large surface area and high electrical conductivity. The RBP antigen and Ru-Nafion@SiO2-labeled secondary antibody were then successively conjugated to form sandwich-type immunocomplexes through the specific interaction between antigen and antibody. The ECL signal amplification was significantly improved due to the synergistic effect of MWCNTs and mesoporous silica nanospheres (mSiO2). The developed ECL immunosensor exhibited high sensitivity and specificity for the detection of RBP and responded linearly to the clinically-relevant concentration of RBP from 78 to 5000 ng mL(-1). Moreover, the MWCNT-based ECL immunosensor displayed excellent stability and reproducibility, as well as successfully achieved the detection of RBP in patient urine samples with desirable results. The present work provided a promising technique for the clinical screening of RBP and point-of-care diagnostics.

A novel sandwich electrochemiluminescence immunosensor for ultrasensitive detection of carbohydrate antigen 19-9 based on immobilizing luminol on Ag@BSA core/shell microspheres.

A novel sandwich-type electrochemiluminescence immunosensor based on immobilizing luminol on Ag@BSA core/shell microspheres (Ag@BSA-luminol) for ultrasensitive detection of tumor marker carbohydrate antigen 19-9 (CA19-9) has been developed. Herein, magnetic carbon nanotubes (MAGCNTs) decorated with polyethylenimine (PEI) was used to construct the base of the immunosensor. MAGCNTs with prominent electrical conductivity and high surface area could be beneficial for promoting the electron transfer and loading plenty of primary antibodies (Ab1) via glutaraldehyde (GA). Meanwhile, the magnetic property of MAGCNTs makes it easy to be attached to the surface of magnetic glass carbon electrode (MGCE) through magnetism interaction, which provides an outstanding platform for this immunosensor. Moreover, Ag@BSA microspheres with large surface area, good stability, and excellent biocompatibility were desirable candidates for effective cross-link of CA19-9 detection antibodies (Ab2). A more interesting thing was that ELISA color reaction was used as an ultrasensitive strategy for identifying Ab2 was successfully coated on Ag@BSA with the naked eye. Additionally, we immobilized the luminol on the surface of Ag@BSA to prepare the target immunosensor. Immobilization of luminol on the surface of Ag@BSA could decrease the distance between luminophores and the electrode surface, leading to great enhancement of the ECL intensity of luminol in the present of hydrogen peroxide (H2O2). Under the optimal conditions, the intensity of the ECL immunosensor increased linearly with the logarithm of CA19-9 concentration in a wide linear range from 0.0005 to 150UmL(-1) with a detection limit of 0.0002UmL(-1) (S/N=3). All the results suggested the prepared CA19-9 immunosensor displayed high sensitivity, excellent stability and good specificity. The developed method opened a new avenue to clinical bioassay.

Electrochemical non-enzyme sensor for detecting clenbuterol (CLB) based on MoS2-Au-PEI-hemin layered nanocomposites

Clenbuterol (CLB), a kind of β2-adrenergic agonists, could disrupt cardiovascular and central nervous system. In this work, a new electrochemical non-enzyme sensor for detecting Clenbuterol (CLB) was fabricated based on MoS2-Au-PEI-hemin layered nanocomposites. The two-dimensional (2-D) MoS2 nanosheets were first in-situ assembled with Au nanoparticles, and polyethylenimine (PEI), then hemin molecules were immobilized onto the MoS2-Au-PEI film-modified glassy carbon electrode (GCE) via amide bond. Scanning electron microscopy (SEM) and Zeta potential measurements were employed to characterize the MoS2-based nanomaterials. Cyclic voltammetry (CV) was used to investigate electrochemical activity of the immobilized hemin on the modified electrode. Upon the optimum conditions, the proposed electrochemical sensor showed an excellent response for CLB including a wide linear ranging from 10ng/mL to 2μg/mL and a detection limit (LOD) of 1.92ng/mL CLB (S/N=3) with favorable reproducibility and stability. Furthermore, this presented method could be feasible for determining CLB in the real pork samples.

Multifunctional nanocarrier based on clay nanotubes for efficient intracellular siRNA delivery and gene silencing

RNA interference-mediated gene silencing relating to disease has recently emerged as a powerful method in gene therapy. Despite the promises, effective transport of siRNA with minimal side effects remains a challenge. Halloysites are cheap and naturally available aluminosilicate clay nanotubes with high mechanical strength and biocompatibility. In this study, a novel multifunctional nanocarrier based on functionalized halloysite nanotubes (f-HNTs) has been developed via electrostatic layer-by-layer assembling approach for loading and intracellular delivery of therapeutic antisurvivin siRNA and simultaneously tracking their intracellular transport, in which PEI-modified HNTs are used as gene vector, antisurvivin siRNA as gene therapeutic agent, and mercaptoacetic acid-capped CdSe quantum dots as fluorescent labeling probes. The successful assembly of the f-HNTs-siRNA complexes was systematically characterized by transmission electron microscopy (TEM), UV–visible spectrophotometry, Zeta potential measurement, fluorescence spectrophotometry, and electrochemical impedance spectroscopy. Confocal microscopy, biological TEM, and flow cytometry studies revealed that the complexes enabled the efficient intracellular delivery of siRNA for cell-specific gene silencing. MTT assays exhibited that the complexes can enhance antitumor activity. Furthermore, Western blot analysis showed that f-HNTs-mediated siRNA delivery effectively knocked down gene expression of survivin and thereby decreased the levels of target proteins of PANC-1 cells. Therefore, this study suggested that the synthesized f-HNTs were a new effective drug delivery system for potential application in cancer gene therapy.

One-step highly sensitive detection of melamine using gold nanoparticle-based dynamic light scattering

In this paper, a dynamic light scattering (DLS)-based approach was developed for the real-time detection of melamine. Due to the strong interaction between melamine and AuNPs, the hydrodynamic diameter of AuNPs changed with increasing amounts of added melamine in the detection system, which can be directly observed by DLS. The sensitivity of this assay to detect melamine is about 0.05 ppm, and even in detection solutions with low pH values (2.5–3.5 or 4.0–5.0), this DLS-based assay can work well. All these results suggest that the DLS-based assay could be a good alternative method for one-step real-time detection of melamine, especially in complex systems (such as complex solutions with a low pH environment), without time-consuming procedures and costly instruments.

A Functional CT Contrast Agent for In Vivo Imaging of Tumor Hypoxia

Hypoxia, which has been well established as a key feature of the tumor microenvironment, significantly influences tumor behavior and treatment response. Therefore, imaging for tumor hypoxia in vivo is warranted. Although some imaging modalities for detecting tumor hypoxia have been developed, such as magnetic resonance imaging, positron emission tomography, and optical imaging, these technologies still have their own specific limitations. As computed tomography (CT) is one of the most useful imaging tools in terms of availability, efficiency, and convenience, the feasibility of using a hypoxia-sensitive nanoprobe (Au@BSA-NHA) for CT imaging of tumor hypoxia is investigated, with emphasis on identifying different levels of hypoxia in two xenografts. The nanoprobe is composed of Au nanoparticles and nitroimidazole moiety which can be electively reduced by nitroreductase under hypoxic condition. In vitro, Au@BSA-NHA attain the higher cellular uptake under hypoxic condition. Attractively, after in vivo administration, Au@BSA-NHA can not only monitor the tumor hypoxic environment with CT enhancement but also detect the hypoxic status by the degree of enhancement in two xenograft tumors with different hypoxic levels. The results demonstrate that Au@BSA-NHA may potentially be used as a sensitive CT imaging agent for detecting tumor hypoxia.

An ESIPT based fluorescent probe for highly selective and ratiometric detection of periodate

Periodate is widely used in organic and bioorganic chemistry, and also related to food and environmental safety. To best of our knowledge, there is no efficient tools reported for simultaneously quantifying periodate with high accuracy and discriminating periodate from other forms of iodine. We have synthesized, characterized and applied a first ratiometric fluorescent probe (PDS-2) for simultaneous monitoring of changes of periodate based on the excited-state intramolecular proton transfer mechanism. This PDS-2 based fluorescent technique may enable for a better understanding of periodate related biological and chemical processes. Also, it is an efficient tool for public health, food safety and environmental protection.

Sensitive detection of tumor cells by a new cytosensor with 3D-MWCNTs array based on vicinal-dithiol-containing proteins (VDPs).

Vicinal-dithiol-containing Proteins (VDPs) are overexpressed in cancer cells and become a potential biomarker for aggressive tumors, the synthesized 2-p-aminophenyl-1, 3, 2-dithiarsenolane (VTA2) is proved to be a highly selective ligand for vicinal dithiols of VDPs in cells. In this report, we developed a new cytosensor based on three-dimensional (3D)-like VTA2-conjugated multiwalled carbon nanotubes (VTA2@MWCNTs) array modified indium tin oxide (ITO) electrode for sensitive and selective detection of VDPs-overexpression tumor cells. The layer-by-layer assembling and cellular detection performances of the 3D-VTA2@MWCNTs-based cytosensor were investigated by atomic force microscopy (AFM) and electrochemical methods including cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS) and differential pulse voltammetry (DPV). Attributed to signal amplification and targeting recognition of the 3D-structured MWCNTs@VTA2, the fabricated cytosensor showed high specificity and sensitivity to the detection of VDPs-overexpression HL-60 cells ranging from 2.7×10(2) to 2.7×10(7)cellsmL(-1) with a low detection limit of ca. 90cellsmL(-1). Furthermore, the captured cancer cells on the cytosensor also could be directly visualized by optical microscopy technology. This proposed 3D-MWCNTs array-based cytosensing strategy provides a simple, sensitive approach for non-invasive cell detection, presenting potential applications in cancer diagnosis.

Electrochemiluminescence resonance energy transfer system between GNRs and Ru(bpy)32+: Application in magnetic aptasensor for β-amyloid

Electrochemiluminescent (ECL) assay has gradually drawn increasing interest in the biomedical analysis. This paper proposed a new methodology for ultrasensitive and facile detection of Alzheimers disease marker β-amyloid (Aβ) by fabricating a sandwich-type ECL sensing platform. Herein, electrochemiluminescence resonance energy transfer (ECL-RET) was employed to determine Aβ concentration, which can be attributed to the quenching effect from RET between Ru(bpy)32+ and gold nanorods (GNRs) acting as ECL-RET electron donor and acceptor, respectively. In this protocol, mesoporous carbon nanospheres were adopted to immobilize ECL reactant Ru(bpy)32+ and antibody via nafion to acquire the RET donor nanocomposites (MOCs/nafion/Ru(bpy)32+/antibody), which were tightly interconnected with epoxy group functionalized Fe3O4 nanoparticles. It is of vital importance that GNRs with exquisite rod shape were synthesized and exhibited a typical absorption peak at 650nm to quench ECL signal of Ru(bpy)32+ effectively. In addition, the ECL emission decreased linearly with the logarithm of Aβ concentration in a wide linear range from 1.0 × 10-5 to 100ng/mL with a detection limit of 4.2 × 10-6ng/mL. Furthermore, distinctive and desirable properties were verified to declare the promise for being applicable to analyze the Aβ content in real Alzheimers cerebrospinal fluid samples with satisfactory results.