Chiral Transport in Nanochannel Based Artificial Drug Transporters.

The precise regulation of chiral drug transmembrane transport can be achieved through drug transporters in living organisms. However, implementing this process in vitro is still a formidable challenge due to the complexity of the biological systems that control drug enantiomeric transport. Herein, a facile and feasible strategy is employed to construct chiral L-tyrosine-modified nanochannels (L-Tyr nanochannels) based on polyethylene terephthalate film, which could enhance the chiral recognition of propranolol isomers (R-/S-PPL) for transmembrane transport. Moreover, conventional fluorescence spectroscopy, patch-clamp technology, laser scanning confocal microscopy, and picoammeter technology are employed to evaluate the performance of nanochannels. The results show that the L-Tyr nanochannel have better chiral selectivity for R-/S-PPL compared with the L-tryptophan (L-Trp) channel, and the chiral selectivity coefficient is improved by about 4.21-fold. Finally, a detailed theoretical analysis of the chirality selectivity mechanism is carried out. The findings would not only enrich the basic theory research related to chiral drug transmembrane transport, but also provide a new idea for constructing artificial channels to separate chiral drugs.

Peptide Nucleic Acid-Functionalized Nanochannel Biosensor for the Highly Sensitive Detection of Tumor Exosomal MicroRNA.

Compared with free miRNAs in blood, miRNAs in exosomes have higher abundance and stability. Therefore, miRNAs in exosomes can be regarded as an ideal tumor marker for early cancer diagnosis. Here, a peptide nucleic acid (PNA)-functionalized nanochannel biosensor for the ultrasensitive and specific detection of tumor exosomal miRNAs is proposed. After PNA was covalently bound to the inner surface of the nanochannels, the detection of tumor exosomal miRNAs was achieved by the charge changes on the surface of nanochannels before and after hybridization (PNA-miRNA). Due to the neutral characteristics of PNA, the efficiency of PNA-miRNA hybridization was improved by significantly reducing the background signal. This biosensor could not only specifically distinguish target miRNA-10b from single-base mismatched miRNA but also achieve a detection limit as low as 75 aM. Moreover, the biosensor was further used to detect exosomal miRNA-10b derived from pancreatic cancer cells and normal pancreatic cells. The results indicate that this biosensor could effectively distinguish pancreatic cancer tumor-derived exosomes from the normal control group, and the detection results show good consistency with those of the quantitative reverse-transcription polymerase chain reaction method. In addition, the biosensor was used to detect exosomal miRNA-10b in clinical plasma samples, and it was found that the content of exosomal miRNA-10b in cancer patients was generally higher than that of healthy individuals, proving that the method is expected to be applied for the early diagnosis of cancer.

Engineering a NO-Regulated Nanofluidic Sensor through the Cyclization Reaction Strategy.

Nitric oxide (NO) is known to be a secondary in vivo signaling agent, demonstrating various biological functions through regulating ion flux in channels. Considering the crucial role of NO in vivo, herein, a biomimetic NO-regulated nanofluidic sensor has been fabricated through a cyclization reaction strategy. This nanofluidic sensor exhibited a promising NO selectivity, sensitivity, and non-interference performance in complex matrices. Thus, such a NO-driven nanosensor will be meaningful for scientific researchers to grasp the in vivo functions of NO.

Switchable Nanochannel Biosensor for H2S Detection Based on an Azide Reduction Reaction Controlled BSA Aggregation.

Hydrogen sulfide (H2S) is the third endogenous gaseous signal molecule in organisms that can directly increase the salt tolerance of plants by closing the K+ channel. Inspired by the important role of H2S in nature, we have designed a H2S-responsive artificial nanochannel biosensor based on an azide reduction reaction strategy. This biomimetic device demonstrates an excellent H2S selective response owing to specific azide reduction by H2S inducing BSA aggregation on the channel with a high ion gating ratio. Furthermore, this H2S-responsive biosensor shows excellent reversibility and stability and a fast response rate, which will help us better understand the synergistic effect of H2S messengers in the ion channels of living organisms.

Ultrasensitive Detection of MicroRNAs with Morpholino-Functionalized Nanochannel Biosensor.

Here, we demonstrate a phosphorodiamidate morpholino oligos (PMO)-functionalized nanochannel biosensor for label-free detection of microRNAs (miRNAs) with ultrasensitivity and high sequence specificity. PMO, as a capture probe, was covalently anchored on the nanochannel surface. Because of the neutral character and high sequence-specific affinity of PMO, hybridization efficiency between PMO and miRNAs was enhanced, thus largely decreasing background signals and highly improving the detection specificity and sensitivity. The miRNAs detection was realized through observing the change of surface charge density when PMO/miRNAs hybridization occurred. Not only could the developed biosensor specifically discriminate complementary miRNAs (Let-7b) from noncomplementary miRNAs (miR-21) and one-base mismatched miRNAs (Let-7c), but also it could detect target miRNAs in serum samples. In addition, this nanochannel-based biosensor attained a reliable limit of detection down to 1 fM in PBS and 10 fM in serum sample, respectively. It is expected that such a new method will benefit miRNA detection in clinical diagnosis.

A surface acoustic wave biosensor synergizing DNA-mediated in situ silver nanoparticle growth for a highly specific and signal-amplified nucleic acid assay.

This work reports a surface acoustic wave (SAW) DNA sensor that synergizes the surface mass effect for signal-amplified and sequence-specific DNA detection in blood serum. By combining an enzyme-mediated DNA extension reaction (both viscoelastic and mass fractions) with the in situ synthesis of silver nanoparticles (mass fraction), a highly sensitive SAW biosensing interface with synergistic mass loading was tailor-engineered. As target DNA hybridized with the surface-confined capture probes, the exposed 3'-OH terminal of the target sequence could be triggered to elongate in the presence of terminal deoxynucleoside transferase (TdT) and deoxy-ribonucleoside triphosphate (dNTP), thereby producing an evident mass effect. Importantly, the extended domain can serve as a template to specifically hybridize with Ag+-binding sequences. In the presence of reducing agents, the accumulated silver ions would nucleate for the in situ synthesis of silver nanoparticles, further enhancing the mass loading. By using this approach, we observed a rapid growth event of silver nanoparticles for signal enhancement, which improved the detection limit (0.8 pM) of the SAW sensor by 3 orders of magnitude as compared to the strategy without signal amplification (at the nanomolar level). The sensor also achieved a high specificity in discriminating even a single-mismatched DNA sequence, and meanwhile could probe the low-abundance DNA molecules directly in human serum with minimal interference. These advantages make the SAW biosensor promising for practical applications, such as monitoring of molecular interactions and disease diagnostics.

Zn(2+) and EDTA Cooperative Switchable Nanofluidic Diode Based on Asymmetric Modification of Single Nanochannel.

Design and fabrication of smart switchable nanofluidic diodes remains a challenge in the life and materials sciences. Here, we present the first example of a novel Zn(2+)/EDTA switchable nanofluidic diode system based on the control of one-side of the modified hourglass-shaped nanochannel with salicylaldehyde Schiff base (SASB). The nanofluidic diode can be turned on in the response of Zn(2+) and turned off in response to EDTA solution with good reversibility and recyclability.

Design and fabrication of a biomimetic nanochannel for highly sensitive arginine response in serum samples.

Inspired from their biological counterparts, chemical modification of the interior surface of nanochannels with functional molecules may provide a highly efficient means to control ionic or molecular transport through nanochannels. Herein, we have designed and prepared a aldehyde calix[4]arene (C4AH), which was attached to the interior surface of a single nanochannel by using a click reaction, and that showed a high response for arginine (Arg). Furthermore, the nanofluidic sensing system has been challenged with complex matrices containing a high concentration of interfering sequences and serum. Based on this finding, we believe that the artificial nanochannel can be used for practical Arg-sensing devices, and be applied in a biological environment.

pH-Stimulated Response Gating for Mimic Cytochrome C Transport on Biomimetic Asymmetric Nanochannels.

Proteins are vital components in cells, biological tissues, and organs, playing a pivotal role in growth and developmental processes in living organisms. Cytochrome C (Cyt C) is a class of heme proteins found in almost all life and is involved in cellular energy metabolic processes such as respiration, mainly as electron carriers or terminal reductases. It binds cardiolipin in the inner mitochondrial membrane, leading to apoptosis. It is a challenge to design a simple and effective artificial system to mimic the complex Cyt C biological transport process. In this paper, an asymmetric biomimetic pH-driven protein gate is described by introducing arginine (Arg) at one end of an hourglass-shaped nanochannel. The nanochannel shows a sensitive protonation-driven protein gate that can be "off" at pH = 7 and "on" at pH = 2. Further studies show that differences in the binding of Arg and Cyt C at different levels of protonation lead to different switching behaviors within the nanochannels, which in turn lead to different surface charges within the nanochannels. It can be used for detecting Cyt C and as an excellent and robust gate for developing integrated circuits and nanoelectronic logic devices.

DSN signal amplification strategy based nanochannels biosensor for the detection of miRNAs.

MiRNA-21 is recognized as an important biological marker for the diagnosis, treatment, and prognosis of breast cancer. Here, we have created a nanochannel biosensor utilizing the duplex-specific nuclease (DSN) signal amplification strategy to achieve the detection of miRNAs. In this system, DNA as the capture probe was covalently immobilized on the surface of nanochannels, which hybridized with the target miRNA and forms RNA/DNA duplexes. DSN could cleave the probe DNA in RNA/DNA duplexes, recycling target miRNA, which may again hybridized with other DNA probes. After N cycles, most of the DNA probes had been cleaved, and the content of miRNA could be quantified by detecting changes in surface charge density. This biosensor can distinguish miR-21 from non-complementary miRNAs and one-base mismatched miRNAs, with reliable detection limits as low as 1 fM in PBS. In addition, we had successfully applied this method to analysis of total RNA samples in MCF-7 cells and HeLa cells, and the nanochannels had also shown excellent responsiveness and strong anti-interference ability. This new method is expected to contribute to miRNA detection in clinical diagnostics, providing a unique approach to detecting and distinguishing disease-associated molecules.

Fabrication of layer-by-layer assembled biomimetic nanochannels for highly sensitive acetylcholine sensing.

Channel tunneling: We have prepared functional biomimetic nanochannels in polyethylene terephthalate (PET) polymer films (see illustration). We used p-sulfonatocalix[4]arene to modify the channel surface by flexible layer-by-layer electrostatic assembly. Using this method we were able to detect acetylcholine with high sensitivity.

Biomimetic ion nanochannels as a highly selective sequential sensor for zinc ions followed by phosphate anions.

A novel biomimetic ion-responsive multi-nanochannel system is constructed by covalently immobilizing a metal-chelating ligand, 2,2'-dipicolylamine (DPA), in polyporous nanochannels prepared in a polymeric membrane. The DPA-modified multi-nanochannels show specific recognition of zinc ions over other common metal ions, and the zinc-ion-chelated nanochannels can be used as secondary sensors for HPO4(2-) anions. The immobilized DPA molecules act as specific-receptor binding sites for zinc ions, which leads to the highly selective zinc-ion response through monitoring of ionic current signatures. The chelated zinc ions can be used as secondary recognition elements for the capture of HPO4(2-) anions, thereby fabricating a sensing nanodevice for HPO4(2-) anions. The success of the DPA immobilization and ion-responsive events is confirmed by measurement of the X-ray photoelectron spectroscopy (XPS), contact angle (CA), and current-voltage (I-V) characteristics of the systems. The proposed nanochannel sensing devices display remarkable specificity, high sensitivity, and wide dynamic range. In addition, control experiments performed in complex matrices suggest that this sensing system has great potential applications in chemical sensing, biotechnology, and many other fields.

Integrated FET sensing microsystem for specific detection of pancreatic cancer exosomal miRNA10b.

Tumor-derived exosome (TD-Ex) serves as a crucial early diagnostic biomarker of pancreatic cancer (PC). However, accurate identification of TD-Ex from PC is still a challenging work. In this paper, a detection microsystem that integrates magnetic separation and FET biosensor is developed, which is capable of selectively separating TD-Ex of PC from the plasma and detecting exosomal miRNA10b in a sensitive and specific manner. The magnetic beads were functionalized with dual antibody (GPC-1 antibody and EpCAM antibody), enabling selective recognition and capture of PC-derived exosomes. On the other hand, a peptide nucleic acid (PNA)- functionalized reduced graphene oxide field-effect transistor (RGO FET) biosensor was subsequently utilized to detect the exosomal miRNA10b, which is highly expressed in PC- derived exosomes. This system could achieve a low detection limit down to 78 fM, and selectively identify miRNA10b from single-base mismatched miRNA. In addition, 40 clinical plasma samples were tested with this microsystem, and the results indicate that it could effectively distinguish PC patients from healthy individuals. The assay combines specific capture and enrichment of PC-derived exosomes with sensitive and selective detection of exosomal miRNA, showing its potential to be used as an effective scheme for PC early diagnosis.

Specific extraction of nucleic acids employing pillar[6]arene-functionalized nanochannel platforms.

The rapid extraction of high-purity nucleic acids from complex biological samples using conventional methods is complicated. Therefore, in this study, glycine-pillar[6]arene (Gly-P6)-functionalized tapered nanochannels were constructed using 32-mer single-stranded E. coli DNA (ssDNA) as a model sequence, which can selectively transport ssDNA by multiple noncovalent forces (transport flux of 2.65 nM cm-2 h-1) under the interference of amino acids and other substances. In view of these prospective results, the selective transport of nucleic acids with nanochannels could be applied in the design of nucleic acid enrichment and separation systems in the future.

Quinolino-triazole linked gold nanoparticles as sensitive 'turn-on' fluorescent Cd(2+) probes.

Quinoline derivatives were brought into the surface of gold nanoparticles (Au NPs) through click chemistry. The fluorescence was quenched by Au NPs because of electron transfer between Au NPs and quinoline. However, upon addition of Cd(2+) to the quinoline-triazole Au NP solution, it exhibited an effective switch-on fluorescence response, owing to the coordination between quinoline and Cd(2+) which can efficiently block the electron transfer. What's more, the fluorescent sensor can effectively detect Cd(2+) in aqueous solution with a detection limit of 1.0 × 10(-5) M.

Nanochannel-based sensor for the detection of lead ions in traditional Chinese medicine.

Lead ions (Pb2+) are used in the quality control of traditional Chinese medicine (TCM) preparations because they are highly toxic to human health. At present, sophisticated analytical instrumentation and complicated procedures for sample analysis are needed for the determination of Pb2+. Herein, a simple, fast, and sensitive peptide-modified nanochannel sensor to detect Pb2+ in TCM is reported, which is based on a Pb2+-specific peptide modified porous anodized aluminum membrane (PAAM). This peptide-based nanochannel clearly has the highest selectivity for Pb2+ when compared to other heavy metal ions, including As2+, Cd3+, Co2+, Cr2+, Cu2+, Fe3+, Hg2+, Mg2+, Mn2+, Ni2+, and Zn2+. Based on linear ranges from 0.01 to 0.16 µM and 10 to 100 µM, the detection limit was calculated to be 0.005 µM. Moreover, this peptide-based nanochannel sensor was successfully used to detect Pb2+ in complex TCM samples. In addition, when compared with the gold standard atomic absorption spectrophotometry (AAS) method, the recovery of the peptide-modified nanochannel sensor was between 87.7% and 116.8%. The experimental results prove that this new sensor is able to achieve accurate detection of Pb2+ in TCM samples. Thus, this sensor system could provide a simple assay for sensitive and selective detection of Pb2+ in TCM, thereby showing great potential in the practical application for the quality control of heavy metals in TCM.

Design and discovery of 1,3-benzodiazepines as novel dopamine antagonists.

A series of novel 1,3-benzodiazapine based D1 antagonists was designed according to the understanding of pharmacophore models derived from SCH 23390 (1b), a potent and selective D1 antagonist. The new design features an achiral cyclic-amidine that maintains desired basicity. Solid phase synthesis was developed for SAR development of the novel dopamine antagonists.

Discovery and SAR of cyclic isothioureas as novel NPY Y1 receptor antagonists.

A class of novel 2-aminobenzothiazoles have been identified as NPY Y(1) antagonists. Various N-heterocyclic substituted aminophenethyl-2-aminobenzothiazole analogs were synthesized to explore the SAR. Isothiourea analogs and ligands with high potency (K(i) 30 nM) have been identified.

Biomimetic nanochannels based biosensor for ultrasensitive and label-free detection of nucleic acids.

A very simple sensing device based on biomimetic nanochannels has been developed for label-free, ultrasensitive and highly sequence-specific detection of DNA. Probe DNA was modified on the inner wall of the nanochannel surface by layer-by-layer (LBL) assembly. After probe DNA immobilization, DNA detection was realized by monitoring the rectified ion current when hybridization occurred. Due to three dimensional (3D) nanoscale environment of the nanochannel, this special geometry dramatically increased the surface area of the nanochannel for immobilization of probe molecules on the inner-surface and enlarged contact area between probes and target-molecules. Thus, the unique sensor reached a reliable detection limit of 10 fM for target DNA. In addition, this DNA sensor could discriminate complementary DNA (c-DNA) from non-complementary DNA (nc-DNA), two-base mismatched DNA (2bm-DNA) and one-base mismatched DNA (1bm-DNA) with high specificity. Moreover, the nanochannel-based biosensor was also able to detect target DNA even in an interfering environment and serum samples. This approach will provide a novel biosensing platform for detection and discrimination of disease-related molecular targets and unknown sequence DNA.

Fabrication of Ultrasensitive Field-Effect Transistor DNA Biosensors by a Directional Transfer Technique Based on CVD-Grown Graphene.

Most graphene field-effect transistor (G-FET) biosensors are fabricated through a routine process, in which graphene is transferred onto a Si/SiO2 substrate and then devices are subsequently produced by micromanufacture processes. However, such a fabrication approach can introduce contamination onto the graphene surface during the lithographic process, resulting in interference for the subsequent biosensing. In this work, we have developed a novel directional transfer technique to fabricate G-FET biosensors based on chemical-vapor-deposition- (CVD-) grown single-layer graphene (SLG) and applied this biosensor for the sensitive detection of DNA. A FET device with six individual array sensors was first fabricated, and SLG obtained by the CVD-growth method was transferred onto the sensor surface in a directional manner. Afterward, peptide nucleic acid (PNA) was covalently immobilized on the graphene surface, and DNA detection was realized by applying specific target DNA to the PNA-functionalized G-FET biosensor. The developed G-FET biosensor was able to detect target DNA at concentrations as low as 10 fM, which is 1 order of magnitude lower than those reported in a previous work. In addition, the biosensor was capable of distinguishing the complementary DNA from one-base-mismatched DNA and noncomplementary DNA. The directional transfer technique for the fabrication of G-FET biosensors is simple, and the as-constructed G-FET DNA biosensor shows ultrasensitivity and high specificity, indicating its potential application in disease diagnostics as a point-of-care tool.