A Sensitive Electrochemiluminescence Urea Sensor for Dynamic Monitoring of Urea Transport in Living Cells.

A multiple signal-amplified electrochemiluminescence (ECL) urea sensor was designed based on a self-enhanced probe and SiO2 photonic crystals for dynamic tracking of urea transmembrane transport. The self-enhanced probe (AuNR@Ru-LA) prepared by loading polyethyleneimine (PEI), lactobionic acid (LA), and Ru(dcbpy)32+ on gold nanorods (AuNRs) generated an initial ECL signal, and then the intensity was multiple-amplified by the enhanced light-scattering effect of SiO2 photonic crystals and the co-reaction with urea. The as-prepared sensor exhibited excellent performance for the detection of urea in the range of 1.0 × 10-10 to 1.0 × 10-4 M with a detection limit of 8.8 × 10-11 M at (3σ)/S. The AuNR@Ru-LA probes were labeled on HepG2 cells to construct a cytosensor with a detection range of 1.0 × 103 to 2.0 × 106 cells mL-1. In addition, the dynamic changes of the extracellular urea concentration were tracked by monitoring the ECL signal of the cytosensor to study urea transmembrane transport. The developed strategy realized the amplification of multiple ECL signals and the tracking of urea transmembrane transport, which provided a novel dynamic detection method for small biomolecules.

Ratiometric ECL sensor based on Apt-AuNS@Lu nanoprobe for analyzing cell swelling-induced ATP release.

A novel ratiometric electrochemiluminescence (ECL) system based on gold nanostars (AuNSs) support was constructed for the determination of hypotonicity-induced ATP release from HepG2 cells. AuNS@Lu nanoprobe was used as anodic luminophore and K2S2O8 as cathodic luminophore as well as anodic co-reactant. AuNS with the large specific surface was adopted to adsorb plentiful luminol to form solid-state probe and as affinity support to immobilize ATP aptamer (Apt). The obtained nanocomposite (Apt-AuNS@Lu) generated a strong ECL signal at + 0.4 V (vs. Ag/AgCl) with co-reactant K2S2O8, because of excellent conductivity and catalytic activity of AuNS. Furthermore, graphene oxide was reduced onto indium tin oxide (ITO) electrodes to facilitate the electron transfer. Following, polydopamine (PDA) film was formed via self-polymerization, improving stability and adhesion of the electrode surface. To immobilize ATP capture aptamer (AptC), abounding AuNSs were attached to RGO/PDA surface. When the sensor was incubated in the mixture solution of Apt-AuNS@Lu and target ATP, the ECL signal of Apt-AuNS@Lu increased with the increase of ATP concentration, meanwhile, the signal of K2S2O8 declined. The ratio of the two luminophores was used for the quantitative determination of ATP. The linear range was 5 to 250 nM, and the limit of detection was 1.4 nM at (3σ)/S. The method was successfully applied to analyze ATP release from HepG2 cells stimulated by 0.45% NaCl hypotonic solution. The results showed that the release kinetics profile of ATP had a sigmoidal shape with rapid release within 10 min and then slowed. Compared to the isotonic groups, the intracellular ATP concentration was 3.7 ± 0.3 µM (n = 3) decreasing by 40.3% and the extracellular was 23.4 ± 1.2 nM (n = 3) increasing by 9.2 times in the hypotonicity for 10 min, which showed ATP release from cells and good agreement with commercial ELISA test. The proposed strategy would be beneficial to broadening application of ECL technology in studying cell biological functions.

A novel signal amplified electrochemiluminescence biosensor based on MIL-53(Al)@CdS QDs and SiO2@AuNPs for trichlorfon detection.

An ultrasensitive electrochemiluminescence (ECL) biosensor was developed based on MIL-53(Al)@CdS QDs and SiO2@AuNPs for trichlorfon detection. Metal-organic frameworks (MOFs) were used as a loading platform that provided a large surface area to load targets and modified materials onto the electrode. At the same time, SiO2@AuNPs loaded plenty of AuNPs which effectively increased the ECL resonance energy transfer between the CdS QDs, so that the ECL signal was strongly quenched and resulted in an amplified response. In the range of 10-11-10-4 M, the ECL response showed a linear relationship with the concentration (logarithm) of trichlorfon, and the detection limit was 5.1 × 10-12 M (S/N = 3). When the biosensor was applied to detect trichlorfon in lettuce, broccoli, cucumber, and chives, the recoveries obtained from the spiked samples were 97%-105%, 102%-104%, 100%-104%, and 98%-104%, respectively. Thus, this novel ECL biosensor has potential applications for the analysis of trichlorfon in food samples.

A photoelectrochemical aptasensor for the sensitive detection of streptomycin based on a TiO2/BiOI/BiOBr heterostructure.

In photoelectrochemical sensor (PEC sensor), sensitivity and selectivity are two essential factors which are determined by photosensitive of materials and identification of elements. Herein, a novel PEC aptamer sensor for streptomycin-specific detection was developed, with which the visible-light-active TiO2/BiOI/BiOBr heterostructure and aptamers were employed as photoactive material and bio-identification elements, separately. The combination of an appropriate amount of TiO2 with BiOI/BiOBr enhanced the photocurrent response, and thus is beneficial to the construction of PEC sensors. In addition, the one-pot synthesis of TiO2/BiOI/BiOBr has the advantage of being environmentally-friendly. Under optimized conditions, the photocurrent response of aptamer/TiO2/BiOI/BiOBr/ITO is linear with SRT concentration from 0.05 to 150 nM, and the detection limit (S/N = 3) is as low as 0.04 nM. This novel PEC sensing strategy provided an ultra-sensitive sensor with high selectivity and stability for SRT detection.

Electrochemiluminescence aptasensor for multiple determination of Hg2+ and Pb2+ ions by using the MIL-53(Al)@CdTe-PEI modified electrode.

An aptasensor based on MIL-53(Al)@CdTe was designed for multiple determination of Hg2+ and Pb2+ by electrochemiluminescence (ECL). Upon the recognition of Hg2+, aptamer 2-AuNPs form hairpin structures and are removed from the electrode. While in the presence of Pb2+, aptamer 1-PtNPs capture the target ions and form G-quadruplexes, and then bring PtNPs close enough to CdTe QDs to produce ECL resonance energy transfer. Upon aptamer interaction with Hg2+ and Pb2+, decreased ECL intensity was observed due to enhanced resonance energy transfer (ERET) and attenuated surface plasmon resonance (SPR). The ECL intensity difference (ΔECL) could therefore be used to detect heavy-metal ions with detection limits of 4.1 × 10-12 M (path 1, Hg2+), 3.7 × 10-11 M (path 2, Pb2+), and 2.4 × 10-11 M (path 3, Pb2+). The aptasensor could also be used for detecting Hg2+ and Pb2+ in fish and shrimp samples with good recoveries.

Electrochemiluminecence nanogears aptasensor based on MIL-53(Fe)@CdS for multiplexed detection of kanamycin and neomycin.

A dual gears electrochemiluminecence (ECL) aptasensing strategy for multiple selective determination of kanamycin and neocycin was designed on the basis of the combination of kanamycin and neocycin induced dual gears conversion, the loading platform of metal-organic frameworks (MOFs), surface plasmon resonance (SPR) and ECL resonance energy transfer (ERET) between CdS QDs and AuNPs (or PtNPs). In the absence of target, the dual gears were "off". Then the B1-AuNP (gear B) and aptamer 1-PtNPs acted as signal quenching elements to quench ECL intensity due to ERET process. Upon addition of kanamycin, the aptamer 1-PtNPs were removed from the gear gradually, the ECL was enhanced due to SPR process between AuNPs and CdS QDs. After the incubation of aptamer 2, the dual gears were "off" again and ECL intensity was decreased by ERET process between AuNPs and CdS QDs. In the presence of neomycin, dual gears were "on" again, the ECL signal was enhanced by SPR process between AuNPs and CdS QDs. Under optimal condition, the proposed aptasensor exhibited wide linear ranges of kanamycin (10-10-10-6 M) and neomycin (10-9-10-5 M), and relatively low detection limits to kanamycin (1.7 × 10-11 M) and neomycin (3.5 × 10-10 M). The developed aptasensor realized the multiple ECL detection of kanamycin and neomycin with single luminophore, and was successfully applied to the detection of kanamycin and neomycin in food samples.

Ru(bpy)32+-Silica@Poly-L-lysine-Au as labels for electrochemiluminescence lysozyme aptasensor based on 3D graphene.

In this work, the feasibility of a novel sensitive electrochemiluminescence aptasensor for the detection of lysozyme using Ru(bpy)32+-Silica@Poly-L-lysine-Au (RuSiNPs@PLL-Au) nanocomposites labeling as an indicator was demonstrated. The substrate electrode of the aptasensor was prepared by depositing gold nanoparticles (AuNPs) on 3D graphene-modified electrode. The lysozyme binding aptamer (LBA) was attached to the 3D graphene/AuNPs electrode through gold-thiol affinity, hybridized with a complementary single-strand DNA (CDNA) of the lysozyme aptamer labeled by RuSiNPs@PLL-Au as an electrochemiluminescence intensity amplifier. Thanks to the synergistic amplification of the 3D graphene, the AuNPs and RuSiNPs@PLL-Au NPs linked to Ru(bpy)32+-ECL further enhanced the ECL intensity of the aptasensor. In presence of lysozyme, the CDNA segment of the self-assembled duplex was displaced by the lysozyme, resulting in decreased electrochemiluminescence signal. Under the optimized conditions, the decrease in electrochemiluminescence intensity varied proportionally with the logarithmic concentration of the lysozyme from 2.25 × 10-12 to 5.0 × 10-8 mol L-1, and the detection limit was estimated to 7.5 × 10-13 mol L-1. The aptasensor was further tested in real samples and found reliable for the detection of lysozyme, thus holding great potential application in food safety researches and bioassay analysis.