Organic Molecular Probe Enabled Ionic Current Rectification toward Subcellular Detection of Glutathione with High Selectivity, Sensitivity, and Recyclability

Liu, Yili; Yu, Siyuan; Chen, Jia-Hao; Wang, Chengshuang; Li, Heng-Ye; Jiang, Dechen; Ye, Deju; Zhao, Weiwei

doi:10.1021/acssensors.2c01897

Cited by 11 publications

(6 citation statements)

References 60 publications

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“…The DNA aptamer could specifically bind the CD63 protein on the exosome surface to reduce the effective pore diameter of the nanopore. The ion current rectification (ICR) effect occurs in asymmetric nanopore structures and was closely related with surface charge and the effective pore diameter. − Hence, binding with the exosome would result in a decrease of ionic current, and the concentration of exosome could be correlated with measured ionic current decrease amplitude. For monitoring of exosomes secretion from single cell, a microwell array chip with small microwell was utilized to capture single cell and the small confined volume (∼390 pL) enables a relative high concentration of secreted exosomes for detection .…”

Section: Resultsmentioning

confidence: 99%

Real-Time Monitoring of Exosomes Secretion from Single Cell Using Dual-Nanopore Biosensors

et al. 2023

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Exosomes secreted from cells carry rich information from their parent cells, representing a promising biomarker for investigation of diseases. We develop a dual-nanopore biosensor using DNA aptamers to specifically recognize CD63 protein on the exosome's surface, which enables label-free exosome detection based on ionic current change. The sensor allows for sensitive detection of exosomes with a detection limit of 3.4 × 10 6 particles/mL. The dual-nanopore biosensor was able to form an intrapipette electric circuit for ionic current measurement due to its unique structure, which is crucial to achieve detection of exosome secretion from a single cell. We utilized a microwell array chip to entrap a single cell into a confined microwell with small volume, enabling the accumulation of exosomes with high concentration. The dual-nanopore biosensor was positioned into the microwell with a single cell, and monitoring of exosome secretion from a single cell in different cell lines and under different stimulations has been achieved. Our design may provide a useful platform for developing nanopore biosensors for detecting cell secretions from a single living cell.

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Section: Resultsmentioning

confidence: 99%

Real-Time Monitoring of Exosomes Secretion from Single Cell Using Dual-Nanopore Biosensors

et al. 2023

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“…As shown in Figure S12, Fe/PSAs-DTSSP displayed a characteristic peak of the thiol group at 2525 cm −1 after being treated with GSH, suggesting the thiol−disulfide exchange reaction between the disulfide bond of Fe/PSAs-DTSSP and GSH to destruct the molecular network. 55,56 Besides, as shown in Figure 3a, Fe/PSAs-DTSSP itself was negatively charged. In the presence of GSH, its thiol group can cleave the disulfide bonds in Fe/PSAs-DTSSP through thiol− disulfide exchange to break down the molecular network, leading the surface of Fe/PSAs-DTSSP to become positively charged.…”

Section: Feasibility Of Highly Specific Colorimetric Detection Of Gsh...mentioning

confidence: 96%

Molecular Network-Bearing Fe–N–C Single-Atom Nanozymes for Monitoring Intracellular Glutathione Fluctuations

Cai,

Chen,

Zhu

et al. 2024

ACS Appl. Nano Mater.

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Nanozymes have drawn much attention owing to their low cost and high stability compared with natural enzymes. However, limited specificity intrinsically presents a challenge to their efficiency in sensing applications. Although specific biorecognition units and molecularly imprinted polymers have been introduced to enhance the specificity of nanozymes, they have some shortcomings like poor stability, high cost, complicated operation, and weakened activity. To address these issues, we developed here a simple and efficient surface engineering strategy to build the target-responsive sensing interface by weaving a molecular network on nanozymes to afford a pocket-like structure. Glutathione (GSH), an important intracellular biothiol, was used as the model target, considering the difficulty in distinguising GSH from other biothiols, especially cysteine (Cys) and homocysteine (Hcy). With high-performance single-atom nanozyme (SAzyme) Fe–N–C as the parent nanozyme, a meshy SAzyme Fe/PSAs-DTSSP containing GSH-cleavable disulfide bonds was fabricated via facile surface modification. Interestingly, the molecular network adhered onto the Fe–N–C surface modulated the peroxidase-like properties, including substrate affinity and catalytic stability under harsh conditions. Moreover, the molecular network could specifically respond to GSH over Cys, Hcy, and other interferents, which suppressed the oxidation of the chromogenic substrate 3,3′,5,5′-tetramethylbenzidine (TMB) due to the reversion of the surface charge property of Fe/PSAs-DTSSP, resulting in a sensitive response identified by the naked eye. The usefulness of this assay was validated by monitoring the fluctuation of the GSH level in cancer cells treated with glucose deprivation or incubated with doxorubicin, showing good accuracy and reliability.

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“…Furthermore, the act of studying a single cell with these analytical techniques typically results in irreversible physiological damage to the cell, which ultimately hinder their development in the field of single cell analysis . In recent years, nanoelectrochemistry has been proved to be a powerful analytical tool for in vivo biomolecule determination due to its high spatial and temporal resolution. − Among them, ionic current rectification (ICR)-based nanopores have been preferred as a promising means for single cell analysis. , Nanopore probes can conduct measuring, sampling, and injecting separately or simultaneously in a tiny space due to their extremely small dimensions. The confined inner space of the nanopore tips avoids the probes from misresponding to external environmental or biological interferences, thus increasing the sensitivity and the specificity of the analytical system.…”

Section: Introductionmentioning

confidence: 99%

Quantification of MicroRNA in a Single Living Cell via Ionic Current Rectification-Based Nanopore for Triple Negative Breast Cancer Diagnosis

Zhang,

Song,

Zheng

et al. 2024

Anal. Chem.

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Accurate analysis of microRNAs (miRNAs) at the single-cell level is extremely important for deeply understanding their multiple and intricate biological functions. Despite some advancements in analyzing singlecell miRNAs, challenges such as intracellular interferences and insufficient detection limits still remain. In this work, an ultrasensitive nanopore sensor for quantitative single-cell miRNA-155 detection is constructed based on ionic current rectification (ICR) coupled with enzyme-free catalytic hairpin assembly (CHA). Benefiting from the enzyme-free CHA amplification strategy, the detection limit of the nanopore sensor for miRNA-155 reaches 10 fM and the nanopore sensor is more adaptable to complex intracellular environments. With the nanopore sensor, the concentration of miRNA-155 in living single cells is quantified to realize the early diagnosis of triplenegative breast cancer (TNBC). Furthermore, the nanopore sensor can be applied in screening anticancer drugs by tracking the expression level of miRNA-155. This work provides an adaptive and universal method for quantitatively analyzing intracellular miRNAs, which will greatly improve our understanding of cell heterogeneity and provide a more reliable scientific basis for exploring major diseases at the single-cell level.

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Organic Molecular Probe Enabled Ionic Current Rectification toward Subcellular Detection of Glutathione with High Selectivity, Sensitivity, and Recyclability

Cited by 11 publications

References 60 publications

Real-Time Monitoring of Exosomes Secretion from Single Cell Using Dual-Nanopore Biosensors

Real-Time Monitoring of Exosomes Secretion from Single Cell Using Dual-Nanopore Biosensors

Molecular Network-Bearing Fe–N–C Single-Atom Nanozymes for Monitoring Intracellular Glutathione Fluctuations

Quantification of MicroRNA in a Single Living Cell via Ionic Current Rectification-Based Nanopore for Triple Negative Breast Cancer Diagnosis

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