Jian Ling

Aptamer-Based Silver Nanoparticles Used for Intracellular Protein Imaging and Single Nanoparticle Spectral Analysis

Aptamer-adapted silver nanoparticles (Apt-AgNPs) were developed as a novel optical probe for simultaneous intracellular protein imaging and single nanoparticle spectral analysis, wherein AgNPs act as an illuminophore and the aptamer as a biomolecule specific recognition unit, respectively. It was found that streptavidin-conjugated and aptamer-functionalized AgNPs show satisfactory biocompatibility and stability in cell culture medium, and thus not only can act as a high contrast imaging agent for both dark-field light scattering microscope and TEM imaging but also can inspire supersensitive single nanoparticle spectra for potential intercellular microenvironment analysis. Further investigations showed that caveolae-related endocytosis is likely a necessary pathway for Apt-AgNPs labeled PrP(c) internalization in human bone marrow neuroblastoma cells (SK-N-SH cells). The integrated capability of Apt-AgNPs to be used as light scattering and TEM imaging agents, along with their potential use for single nanoparticle spectral analysis, makes them a great promise for future biomedical imaging and disease diagnosis.

Sensitive Discrimination and Detection of Prion Disease-Associated Isoform with a Dual-Aptamer Strategy by Developing a Sandwich Structure of Magnetic Microparticles and Quantum Dots

The major challenge of prion disease diagnosis at the presymptomatic stage is how to sensitively or selectively discriminate and detect the minute quantity of disease-associated prion protein isoform (PrP(Res)) in complex biological systems such as serum and brain homogenate. In this contribution, we developed a dual-aptamer strategy by taking the advantages of aptamers, the excellent separation ability of magnetic microparticles (MMPs), and the high fluorescence emission features of quantum dots (QDs). Two aptamers (Apt1 and Apt2), which can recognize their two corresponding distinct epitopes of prion proteins (PrP), were coupled to the surfaces of MMPs and QDs, respectively, to make MMPs-Apt1 and QDs-Apt2 ready at first, which then could be coassociated together through the specific recognitions of the two aptamers with their two corresponding distinct epitopes of PrP, forming a sandwich structure of MMPs-Apt1-PrP-Apt2-QDs and displaying the strong fluorescence of QDs. Owing to the different binding affinities of the two aptamers with PrP(Res) and cellular prion protein (PrP(C)), both of which have distinct denaturing detergent resistance, our dual-aptamer strategy could be applied to discriminate PrP(Res) and PrP(C) successfully in serum. Further identifications showed that the present dual-aptamer assay could be successfully applied to the detection of PrP in 0.01% brain homogenate, about 1000-fold lower than that of commonly applied antibody-mediated assays, which can detect PrP just in 10% brain homogenate, indicating that the present designed dual-aptamer assay is highly sensitive and adequate for clinical diagnosis without isolation of target protein prior to assay.

Visual Sandwich Immunoassay System on the Basis of Plasmon Resonance Scattering Signals of Silver Nanoparticles

In this contribution, we established a sandwich immunoassay system with a common spectrofluorometer to collect the plasmon resonance scattering (PRS) signals from silver nanoparticles (AgNPs) immunotargeted on glass slides. By taking the immunoreactions of goat antihuman IgG (Fc fragment specific) antibody (GAH-IgG), human immunoglobulin (H-IgG), and rabbit antihuman IgG (Fab fragment specific) antibody (RAH-IgG) as an example, we found that if a primary antibody (GAH-IgG) was first immobilized on the surface of glass slides and applied to capture target antigen (H-IgG), AgNPs-labeled secondary antibody (RAH-IgG) could be employed to detect the target antigen (H-IgG) by forming a sandwich immune complex on the surface of the glass slide. It was found that the PRS signals resulting from the AgNPs immunotargeted on the glass slides could be applied to the quantitative detection of H-IgG target antigen in the range of 10-1000 ng/mL with the limit of determination of 1.46 ng/mL (3sigma) under optimal conditions, which is sensitive and comparable with reported chemiluminescence immunoassays. With a dark-field microscope coupled with a spectral system, we measured the PRS features of single AgNPs immunotargeted on the glass slides, showing that the PRS of single nanoparticles might have potential applications in analytical chemistry. Further findings showed that the strong PRS signals from the AgNPs immunotargeted on the glass slides can be clearly seen and distinguished by naked eyes under the excitation of a common white light-emitting diode (LED) torch. Therefore, a visual PRS immunoassay system can be established easily with common glass slides and an LED torch.

Visual colorimetric detection of berberine hydrochloride with silver nanoparticles

A visual colorimetric method for the detection of berberine hydrochloride was proposed in this contribution based on the color change caused by the aggregation of silver nanoparticles (AgNps). It was found that citrate-capped AgNps dispersed in water owing to the electrostatic repulsion from each other by the negative charged surface, presenting a bright yellow color. However, the presence of positively charged berberine could induce the aggregation of citrate-capped AgNps, resulting in color change from yellow to green and even to blue depending on the concentration of berberine. Under the optimum condition investigated with UV-vis absorption and light scattering technique, berberine hydrochloride from 0.05 micromol l(-1) to 0.4 micromol l(-1) could be visually detected based on the color alteration of the AgNps suspension.

Nanotoxicity of Silver Nanoparticles to Red Blood Cells: Size Dependent Adsorption, Uptake, and Hemolytic Activity

Silver nanoparticles (AgNPs) are increasingly being used as antimicrobial agents and drug carriers in biomedical fields. However, toxicological information on their effects on red blood cells (RBCs) and the mechanisms involved remain sparse. In this article, we examined the size dependent nanotoxicity of AgNPs using three different characteristic sizes of 15 nm (AgNPs15), 50 nm (AgNPs50), and 100 nm (AgNPs100) against fish RBCs. Optical microscopy and transmission electron microscopy observations showed that AgNPs exhibited a size effect on their adsorption and uptake by RBCs. The middle sized AgNPs50, compared with the smaller or bigger ones, showed the highest level of adsorption and uptake by the RBCs, suggesting an optimal size of ∼50 nm for passive uptake by RBCs. The toxic effects determined based on the hemolysis, membrane injury, lipid peroxidation, and antioxidant enzyme production were fairly size and dose dependent. In particular, the smallest sized AgNPs15 displayed a greater ability to induce hemolysis and membrane damage than AgNPs50 and AgNPs100. Such cytotoxicity induced by AgNPs should be attributed to the direct interaction of the nanoparticle with the RBCs, resulting in the production of oxidative stress, membrane injury, and subsequently hemolysis. Overall, the results suggest that particle size is a critical factor influencing the interaction between AgNPs and the RBCs.

A colorimetric method for highly sensitive and accurate detection of iodide by finding the critical color in a color change process using silver triangular nanoplates

In this contribution, we demonstrated a novel colorimetric method for highly sensitive and accurate detection of iodide using citrate-stabilized silver triangular nanoplates (silver TNPs). Very lower concentration of iodide can induce an appreciable color change of silver TNPs solution from blue to yellow by fusing of silver TNPs to nanoparticles, as confirmed by UV-vis absorption spectroscopy and transmission electron microscopy (TEM). The principle of this colorimetric assay is not an ordinary colorimetry, but a new colorimetric strategy by finding the critical color in a color change process. With this strategy, 0.1 μM of iodide can be recognized within 30 min by naked-eyes observation, and lower concentration of iodide down to 8.8 nM can be detected using a spectrophotometer. Furthermore, this high sensitive colorimetric assay has good accuracy, stability and reproducibility comparing with other ordinary colorimetry. We believe this new colorimetric method will open up a fresh insight of simple, rapid and reliable detection of iodide and can find its future application in the biochemical analysis or clinical diagnosis.

Energy transfer with gold nanoparticles for analytical applications in the fields of biochemical and pharmaceutical sciences

Gold nanoparticles (AuNPs) are the most interesting nanomaterials for analytical purposes owing to their unique optical and electrochemical properties resulting from the localized surface plasmon resonance (LSPR) of the electrons, displaying strong absorption and scattering of light from visible to near-infrared region by tuning the particles sizes and shapes. In fluorescence resonance energy transfer (FRET) process, AuNPs have been identified to act as excellent acceptors to replace traditional organic quenchers. Starting from quenching advantages of AuNPs in FRET system and energy transfer mechanism of donor to AuNPs, herein we try to summarize and discuss typical AuNPs based FRET methods in terms of DNA hybridizations, immunoreactions, specific molecular bindings or adsorptions, and provide their analytical applications in biochemical and pharmaceutical analysis.

Recent Developments of the Resonance Light Scattering Technique: Technical Evolution, New Probes and Applications

Abstract The resonance light scattering (RLS) technique, scanning simultaneously the excitation and emission monochromators of a common spectrofluorometer to detect enhanced RLS signals, has been used for designating bio‐assemblies, aggregation species, and analytical purposes. Herein, we review the reports since our last mini‐review in 2003 concerning the new derived RLS techniques, RLS probes, and their applications.

Magnetic particle-based sandwich sensor with DNA-modified carbon nanotubes as recognition elements for detection of DNA hybridization.

In this contribution, we design a visual sensor for DNA hybridization with DNA probe-modified magnetic particles (MPs) and multiwalled carbon nanotubes (MWNTs) without involving a visual recognition element such as fluorescent/chemiluminescent reagents. It was found that DNA probe-modified MWNTs, which could be dispersed in aqueous medium and have strong light scattering signals under the excitation of a light beam in the UV-vis region, could connect with DNA probe-modified MPs together in the presence of perfectly complementary target DNA and form a sandwich structure. In a magnetic field, the formed MP-MWNT species can easily be removed from the solution, resulting in a decrease of light scattering signals. Thus, a magnetic particle-based sandwich sensor could be developed to detect DNA hybridization by measuring the light scattering signals with DNA-modified MWNTs as recognition elements. Experiments showed that the DNA-modified MPs sensor could be reused at least 17 times and was stable for more than 6 months.

A label-free visual immunoassay on solid support with silver nanoparticles as plasmon resonance scattering indicator

By taking silver nanoparticles (Ag-NPs) as plasmon resonance scattering (PRS) indicator considering that Ag-NPs have strong plasmon resonance light scattering signals corresponding to their plasmon resonance absorption (PRA), we propose a label-free visual immunoassay on the solid support of glass slides. Our investigations showed that Ag-NPs could be adsorbed on the surface of glass slides where immunoreactions between a previously immobilized antigen and its antibody have occurred if the glass slides were immersed in an Ag-NP suspension whose pH value has been carefully adjusted. The optimal pH of the Ag-NP suspension depends on the nature of previously immobilized antigen and its antibody. It was found that the adsorption of negative-charged Ag-NPs on the surface of glass slides depends only on the content of antibody under optimal conditions. With a common spectrofluorometer to measure the PRS signals of the Ag-NPs adsorbed on the surface, we could detect antibody in the range of 10 to 160 ng ml(-1). If a white light-emitting diode (LED) torch is employed to illuminate the glass slides, we can make visual detection of the antibody by the naked eye.