Jimin Gao

A Graphene-Based Sensor Array for High-Precision and Adaptive Target Identification with Ensemble Aptamers

In this work, we report a new concept of adaptive ensemble aptamers (ENSaptamers) that exploits the collective recognition abilities of a small set of rationally designed, nonspecific DNA sequences to identify molecular or cellular targets discriminatively. In contrast to in vitro-selected aptamers, which possess specific lock-and-key recognition, ENSaptamers rely on pattern recognition that mimics natural olfactory or gustatory systems. Nanographene oxide was employed to provide a low-background and highly reproducible fluorescent assay system. We demonstrate that this platform provides a highly discriminative and adaptive tool for high-precision identification of a wide range of targets for diagnostic and proteomic applications with a nearly unlimited supply of ENSaptamer receptors.

A dumbbell probe-mediated rolling circle amplification strategy for highly sensitive microRNA detection

We herein report the design of a dumbbell-shaped DNA probe that integrates target-binding, amplification and signaling within one multifunctional design. The dumbbell probe can initiate rolling circle amplification (D-RCA) in the presence of specific microRNA (miRNA) targets. This D-RCA-based miRNA strategy allows quantification of miRNA with very low quantity of RNA samples. The femtomolar sensitivity of D-RCA compares favorably with other existing technologies. More significantly, the dynamic range of D-RCA is extremely large, covering eight orders of magnitude. We also demonstrate miRNA quantification with this highly sensitive and inexpensive D-RCA strategy in clinical samples.

Programmable Engineering of a Biosensing Interface with Tetrahedral DNA Nanostructures for Ultrasensitive DNA Detection

Self-assembled DNA nanostructures with precise sizes allow a programmable soft lithography approach to engineer the interface of electrochemical DNA sensors. By using millimeter-sized gold electrodes modified with several types of tetrahedral DNA nanostructures (TDNs) of different sizes, both the kinetics and thermodynamics of DNA hybridization were profoundly affected. Because each DNA probe is anchored on an individual TDN, its lateral spacing and interactions are finely tuned by the TDN size. By simply varying the size of the TDNs, the hybridization time was decreased and the hybridization efficiency was increased. More significantly, the detection limit for DNA detection was tuned over four orders of magnitude with differentially nanostructured electrodes, and achieved attomolar sensitivity with polymeric enzyme amplification.

Nanomaterials-based sensors for applications in environmental monitoring

Nanomaterials are well known to possess excellent electrical, optical, thermal, catalytic properties and strong mechanical strength, which offer great opportunities to construct nanomaterials-based sensors or devices for monitoring environmental contaminations in air, water and soil. Various nanomaterials, such as carbon nanotubes, gold nanoparticles, silicon nanowires and quantum dots, have been extensively explored in detecting and measuring toxic metal ions, toxic gases, pesticides, and hazardous industrial chemicals with high sensitivity, selectivity and simplicity. In the feature article, we reviewed recent advances in this direction, by classifying nanomaterials into five categories to illustrate the applications of nanomaterials in environmental monitoring.

A carbon nanotube-based high-sensitivity electrochemical immunosensor for rapid and portable detection of clenbuterol

Carbon nanotubes have shown their unique advantages of mechanical, chemical and electronic properties in bioanalysis. We herein report a new method to efficiently and reproducibly prepare multi-walled carbon nanotubes (MWNTs)-protein sensing layers for electrochemical immunosensors. This method employs centrifugation to prepare a conjugate of MWNTs and goat anti mouse-immunoglobulin G (IgG) (secondary antibody). The conjugates were then deposited on screen-printed electrodes to form a nanostructured layer (MWNT-I layer). CLB monoclonal antibody was assembled through its binding to the secondary antibody. The MWNT-I layer-based electrodes were used for rapid and sensitive amperometric immunosensing detection of clenbuterol (CLB) in swine urine samples. Horseradish peroxidase-coupled CLB (CLB-HRP) competed with free CLB in the samples to bind the monoclonal antibody. It has shown significantly higher sensitivity and better reproducibility than the chemical conjugation method. This MWNT-based immunosensor is highly sensitive, leading to a limit of detection of 0.1 ng/mL within a rapid assay time of 16 min. Its sensitivity is at least 1 order of magnitude higher than that of a normal immunosensor (without MWNTs). The sensing device is portable with disposable screen-printed electrode, satisfactorily meeting the requirements for field detection of food security-related species.

Ultrasensitive Electrochemical Detection of Prostate-Specific Antigen by Using Antibodies Anchored on a DNA Nanostructural Scaffold

The high occurrence of prostate cancer in men makes the prostate-specific antigen (PSA) screening test really important. More importantly, the recurrence rate after radical prostatectomy is high, whereas the traditional PSA immunoassay does not possess the sufficient high sensitivity for post-treatment PSA detection. In these assays, uncontrolled and random orientation of capture antibodies on the surface largely reduces their activity. Here, by exploiting the rapidly emerging DNA nanotechnology, we developed a DNA nanostructure based scaffold to precisely control the assembly of antibody monolayer. We demonstrated that the detection sensitivity was critically dependent on the nanoscale-spacing (nanospacing) of immobilized antibodies. In addition to the controlled assembly, we further amplified the sensing signal by using the gold nanoparticles, resulting in extremely high sensitivity and a low detection limit of 1 pg/mL. To test the real-world applicability of our nanoengineered electrochemical sensor, we evaluated the performance with 11 patients serum samples and obtained consistent results with the gold-standard assays.

Gold Nanoparticle-Based Enzyme-Linked Antibody-Aptamer Sandwich Assay for Detection of Salmonella Typhimurium

Enzyme-linked immunosorbent assay (ELISA) provides a convenient means for the detection of Salmonella enterica serovar Typhimurium (STM), which is important for rapid diagnosis of foodborne pathogens. However, conventional ELISA is limited by antibody-antigen immunoreactions and suffers from poor sensitivity and tedious sample pretreatment. Therefore, development of novel ELISA remains challenging. Herein, we designed a comprehensive strategy for rapid, sensitive, and quantitative detection of STM with high specificity by gold nanoparticle-based enzyme-linked antibody-aptamer sandwich (nano-ELAAS) method. STM was captured and preconcentrated from samples with aptamer-modified magnetic particles, followed by binding with detector antibodies. Then nanoprobes carrying a large amount of reporter antibodies and horseradish peroxidase molecules were used for colorimetric signal amplification. Under the optimized reaction conditions, the nano-ELAAS assay had a quantitative detection range from 1 × 10(3) to 1 × 10(8) CFU mL(-1), a limit of detection of 1 × 10(3) CFU mL(-1), and a selectivity of >10-fold for STM in samples containing other bacteria at higher concentration with an assay time less than 3 h. In addition, the developed nanoprobes were improved in terms of detection range and/or sensitivity when compared with two commercial enzyme-labeled antibody signal reporters. Finally, the nano-ELAAS method was demonstrated to work well in milk samples, a common source of STM contamination.

Pattern Recognition Analysis of Proteins Using DNA‐Decorated Catalytic Gold Nanoparticles

A label-free protein analysis strategy is based on patterns of gold nanoparticle (AuNP) growth. AuNPs pretreated with different oligonucleotides are challenged with various proteins. After Au reduction, the colorimetric patterns are processed with linear discriminant analysis. This method discriminates different proteins, or one protein of different concentrations, in mixed samples or even serum and urine.

Ultrasensitive electrochemical DNA sensor based on the target induced structural switching and surface-initiated enzymatic polymerization

In this work, two electrochemical DNA sensors was developed based on the target induced structural switching of stem-loop probe (SLP) and surface initiated enzymatic polymerization (SIEP). Both of the electrochemical DNA sensors employed SLPs with the same sequence. However, one had a thiol label at its 3 terminal (the probe was named 3-SLP and the sensor was named 3-SLP-SENS) and the other at its 5 terminal (the probe was named 5-SLP and the sensor was named 5-SLP-SENS). In the initial state of the sensors, both of the probes adopted the stem-loop structure, which shielded the unlabeled terminals of capture probes from being approached. When the loop regions of the capture probes hybridized with the target DNA the conformation of the SLPs was changed to a rigid double-strand, as a result, the 5-SLP released a 3-OH terminal for SIEP which could be catalyzed by terminal deoxynucleotidyl transferase (TdT). And the 3-SLP released a 5 phosphate terminal which is not suit for SIEP. Thus a signal probe was employed to hybridize with the 5 terminal of 3-SLP and provide a 3-OH. Both of the sensors were then submitted to the TdT-mediated SIEP. By using biotinylated 2-deoxyadenosine 5-triphosphate (biotin-dATP), biotin labels are incorporated into the SIEP-generated long single-stranded DNA. Then avidin-horseradish peroxidases (Av-HRPs) were employed for specific binding to the biotin labels to produce electrochemical signals. The detection performances of two electrochemical DNA sensors were investigated and compared. It was demonstrated that though the 3-SLP-SENS employed extra signal probes, the background current was lower leading to a better detection limit. By taking advantage of SLP and SIEP, this 3-SLP-SENS has been able to detect as low as 0.1pM DNA targets with excellent differentiation ability for even single mismatches.

Binding-induced collapse of DNA nano-assembly for naked-eye detection of ATP with plasmonic gold nanoparticles.

The detection of small molecules depends heavily on complicated GC-MS (Gas chromatography-mass spectrometry), HPLC (High-performance liquid chromatography) and some other complicated instruments that are not suitable for point of care detection. Here, we have demonstrated a fast (in 10min), simple (instrument-free) and effective detection platform for small molecule-ATP. In our design, we engineered the hybridization region of aptamer and assembled it into a superstructure to avoid the exposed flexible ends. The binding of ATP triggered the collapse of the superstructures to produce single stranded DNA that can obviously tune the plasmonic coupling of unmodified gold nanoparticles (AuNPs). Compared to detection platforms based on fully hybridized aptamer double helix, the detection time was significantly decreased to 10min. The resulting color change can be recognized by naked eyes. Our detection is highly specific and selective. Furthermore, a logic gate with multiplexed detection capability for ATP and DNA were demonstrated.