An n-Type Open-Shell Conjugated Polymer with High-Spin Ground-State and High Intrinsic Electrical Conductivity.

Open-shell conjugated polymers with a high intrinsic conductivity and high-spin ground state hold considerable promise for applications in organic electronics and spintronics. Herein, two novel acceptor-acceptor (A-A) conjugated polymers based on a highly electron-deficient quinoidal benzodifurandione unit have been developed, namely DPP-BFDO-Th and DPP-BFDO. The incorporation of the quinoidal moiety into the polymers backbones enables deeply aligned lower-lying lowest unoccupied molecular orbital (LUMO) levels of below -4.0 eV. Notably, DPP-BFDO exhibits an exceptionally low LUMO (-4.63 eV) and a high-spin ground state characterized by strong diradical characters. Moreover, a self-doping through intermolecular charge-transfer is observed for DPP-BFDO, as evidenced by X-ray photoelectron spectroscopy (XPS) studies. The high carrier concentration in combination with a planar and linear conjugated backbone yields a remarkable electrical conductivity (σ) of 1.04 S cm-1 in the "undoped" native form, ranking among the highest values reported for n-type radical-based conjugated polymers. When employed as an n-type thermoelectric material, DPP-BFDO achieves a power factor of 12.59 µW m-1 K-2. Furthermore, upon n-doping, the σ could be improved to 65.68 S cm-1. This study underscores the great potential of electron-deficient quinoidal units in constructing dopant-free n-type conductive polymers with a high-spin ground state and exceptional intrinsic conductivity.

Solid-State Glass Nanopipettes: Functionalization and Applications.

Solid-state glass nanopipettes provide a promising confined space that offers several advantages such as controllable size, simple preparation, low cost, good mechanical stability, and good thermal stability. These advantages make them an ideal choice for various applications such as biosensors, DNA sequencing, and drug delivery. In this review, we first delve into the functionalized nanopipettes for sensing various analytes and the methods used to develop detection means with them. Next, we provide an in-depth overview of the advanced functionalization methodologies of nanopipettes based on diversified chemical kinetics. After that, we present the latest state-of-the-art achievements and potential applications in detecting a wide range of targets, including ions, molecules, biological macromolecules, and single cells. We examine the various challenges that arise when working with these targets, as well as the innovative solutions developed to overcome them. The final section offers an in-depth overview of the current development status, newest trends, and application prospects of sensors. Overall, this review provides a comprehensive and detailed analysis of the current state-of-the-art functionalized nanopipette perception sensing and development of detection means and offers valuable insights into the prospects for this exciting field.

Modification of Food Allergy on the Associations between Early Life Exposure to Size-Specific Particulate Matter and Childhood Allergic Rhinitis.

Previous studies have reported the association between particulate matter (PM) and childhood allergic rhinitis (AR). However, it is unclear whether food allergy (FA) modifies the PM-AR association. We aimed at evaluating the effect of the modification of FA on PM-AR association in preschool children. We adopted a cross-sectional study and conducted a questionnaire survey among preschool children aged 3-6 years in 7 cities in China from June 2019 to June 2020 to collect information on AR and FA. We used a combination of multilevel logistic regression and restricted cubic spline functions to quantitatively assess whether FA modifies the associations between size-specific PM exposure (1 × 1 km) and the risk of AR. The adjusted odds ratios (ORs) for AR among the children with FA as per a 10 µg/m3 increase in early life PM1, PM2.5, and PM10 were significantly higher than the corresponding ORs among the children without FA [e.g., OR: 1.58, 95% CI: (1.32, 1.90) vs 1.29, 95% CI: (1.18, 1.41), per 10 µg/m3 increase in PM1]. The interactions between FA and size-specific PM exposure and their effects on AR were statistically significant (all p-int < 0.001). FA, as an important part of the allergic disease progression, may modify the PM-AR association in preschool children.

Discovery, Optimization, and Biological Evaluation of Arylpyridones as Cbl-b Inhibitors.

Casitas B-lymphoma proto-oncogene-b (Cbl-b), a member of the Cbl family of RING finger E3 ubiquitin ligases, has been demonstrated to play a central role in regulating effector T-cell function. Multiple studies using gene-targeting approaches have provided direct evidence that Cbl-b negatively regulates T, B, and NK cell activation via a ubiquitin-mediated protein modulation. Thus, inhibition of Cbl-b ligase activity can lead to immune activation and has therapeutic potential in immuno-oncology. Herein, we describe the discovery and optimization of an arylpyridone series as Cbl-b inhibitors by structure-based drug discovery to afford compound 31. This compound binds to Cbl-b with an IC50 value of 30 nM and induces IL-2 production in T-cells with an EC50 value of 230 nM. Compound 31 also shows robust intracellular target engagement demonstrated through inhibition of Cbl-b autoubiquitination, inhibition of ubiquitin transfer to ZAP70, and the cellular modulation of phosphorylation of a downstream signal within the TCR axis.

Interface Engineering for Highly Efficient Organic Solar Cells.

Organic solar cells (OSCs) have made dramatic advancements during the past decades owing to the innovative material design and device structure optimization, with power conversion efficiencies surpassing 19% and 20% for single-junction and tandem devices, respectively. Interface engineering, by modifying interface properties between different layers for OSCs, has become a vital part to promote the device efficiency. It is essential to elucidate the intrinsic working mechanism of interface layers, as well as the related physical and chemical processes that manipulate device performance and long-term stability. In this article, the advances in interface engineering aimed to pursue high-performance OSCs are reviewed. The specific functions and corresponding design principles of interface layers are summarized first. Then, the anode interface layer, cathode interface layer in single-junction OSCs, and interconnecting layer of tandem devices are discussed in separate categories, and the interface engineering-related improvements on device efficiency and stability are analyzed. Finally, the challenges and prospects associated with application of interface engineering are discussed with the emphasis on large-area, high-performance, and low-cost device manufacturing.

Influence of Interaction between Electrolyte with Side-Chain Free Conjugated Polymer on the Performance of Organic Electrochemical Transistors.

Organic electrochemical transistors (OECTs) offer significant advantages in electrophysiological applications, primarily due to their ability to facilitate ionic-to-electronic conversion and establish a direct interface with the surrounding aqueous environments by using organic mixed ionic-electronic conductors. This study employs a side-chain free n-type conducting polymer, poly(benzodifurandione) (PBFDO), as the channel material in OECTs to scrutinize the interplay between various ion concentrations in electrolytes and the conjugated polymer and to assess their subsequent impact on device performance. Our findings reveal that PBFDO-based OECTs demonstrate superior transfer characteristics, attributed to their high conductivity and remarkable stability in aqueous solutions. Interestingly, the ion concentration does not alter the electronic band structure of PBFDO during the doping process, but a high-salt-concentration electrolyte could accelerate the electrochemical process compared to its counterparts. Furthermore, the diluted solution significantly enhances the surface roughness and decreases the crystalline coherence length of the film compared with concentrated solutions. A quantitative analysis utilizing an electrochemical quartz crystal microbalance discloses that the electrolyte ions penetrate the PBFDO film, inducing the absorption of a fraction of water molecules, which is pronounced in diluted solutions and negligible in their concentrated counterparts. This notable swelling of the polymer in diluted solutions potentially hampers the transport of charge carriers, consequently diminishing the OECT performance. This research elucidates a direct correlation between microstructure alterations and device performance during operation, paving the way for the optimization of ionic and electronic conductivity in polymers to foster the development of high-performance organic electronic devices.

Discovery of a Novel Benzodiazepine Series of Cbl-b Inhibitors for the Enhancement of Antitumor Immunity.

Casitas B-lineage lymphoma proto-oncogene-b (Cbl-b) is a RING finger E3 ligase that is responsible for repressing T-cell, natural killer (NK) cell, and B-cell activation. The robust antitumor activity observed in Cbl-b deficient mice arising from elevated T-cell and NK-cell activity justified our discovery effort toward Cbl-b inhibitors that might show therapeutic promise in immuno-oncology, where activation of the immune system can drive the recognition and killing of cancer cells. We undertook a high-throughput screening campaign followed by structure-enabled optimization to develop a novel benzodiazepine series of potent Cbl-b inhibitors. This series displayed nanomolar levels of biochemical potency, as well as potent T-cell activation. The functional activity of this class of Cbl-b inhibitors was further corroborated with ubiquitin-based cellular assays.

Hairpin DNA-based electrochemical amplification strategy for miRNA sensing by using single gold nanoelectrodes.

A new sensor has been developed to detect miRNA-15 using nanoelectrodes and a hairpin DNA-based electrochemical amplification technique. By utilizing a complex DNA cylinder connected with hairpin DNA1, the sensor is able to absorb more methylene blue (MB) than simple double-stranded DNA. Another hairpin DNA2 is modified on an Au nanoelectrode surface and, when miRNA-15 is introduced, it triggers a chain reaction. This reaction unlocks two hairpins alternatively to polymerize into a complex structure that attaches more MB. The miRNA-15 is then replaced by DNA1 due to strand displacement reactions and continues to react with the next DNA2 to achieve circular amplification. The electrochemical signal from MB oxidation has a linear relationship with the miRNA-15 concentrations, making it possible to detect miRNA-15. Moreover, this method can be readily adapted for the detection of various other miRNA species. The newly devised nanosensor holds promising applications for the in vivo detection of miRNA-15 within biological systems, which is achieved by leveraging the advantageous characteristics of nanoelectrodes, including their low resistance-capacitance time constant, rapid mass transfer kinetics, and small diameter.

Discovery and characterisation of quinazolines and 8-Azaquinazolines as NLRP3 agonists with oral bioavailability in mice.

The NLRP3 inflammasome is a multiprotein complex that plays a critical role in activating the immune system in response to danger signals. Small molecule agonists of NLRP3 may offer clinical benefits in cancer immunology either as a monotherapy or in combination with checkpoint blockade, where it is hypothesised that their application can help to initiate an antitumor immune response. In this study, we report the discovery of quinazolines and 8-azaquinazolines as NLRP3 agonists and their chemical optimization to afford compounds with oral bioavailability in mice. We confirm that these compounds engage the NLRP3 inflammasome by verifying their dependence upon lipopolysaccharide (LPS) priming for cytokine release and the activation of Caspase-1. We further demonstrate pathway engagement through loss of activity in an NLRP3-knockout THP1 cell line. Based on their pharmacokinetic profile and biological activity, these compounds represent valuable tools to evaluate the therapeutic potential of NLRP3 activation in a pre-clinical setting.

Multiplex Assays of MicroRNAs by Using Single Particle Electrochemical Collision in a Single Run.

It is important to quantify multiple biomarkers in a single run due to the advantages of precious samples and diagnostic accuracy. Based on the distinguishability of two types of current signals from single particle electrochemical collision (SPEC), step-type current transients produced by Pt nanoparticles (PtNPs) catalyzed hydrazine oxidation and peak-type current transients produced by Ag nanoparticles (AgNPs) oxidation, a kind of multiplex immunoassay of target microRNAs (miRNA-21 and Let-7a) have been established during SPEC in a single run. When the single-stranded DNA (ssDNA1) that was perfectly complementary to miRNA-21 was coupled to the surface of PtNPs, the SPEC of PtNPs electrocatalysis was inhibited and the step-type current transients disappeared, while the single-stranded DNA (ssDNA2) that was perfectly complementary to Let-7a was coupled to the surface of AgNPs, the SPEC of AgNPs oxidation was inhibited, and the peak-type current transients disappeared, thus the signals were in the "off" state at this time. After that, miRNA-21 and Let-7a were added into solution, complementary base pairing disrupted the weak DNA-NP interaction and restored the electrocatalysis of PtNPs and the electrooxidation of AgNPs, and the step-type current signals and peak-type current signals were in the "on" state. Moreover, the frequencies from two different recovered signals (PtNPs catalysis and AgNPs oxidation) corresponded to the amount of added miRNA-21 and Let-7a, thus a multiplex immunoassay method for dual quantification of miRNA-21 and Let-7a in a single run was established.

Compound probiotics producing cellulase could replace cellulase preparations during solid-state fermentation of millet bran.

Low-value agricultural by-products can be converted into high-value biological products by fermentation with probiotic strains or by enzymatic hydrolysis. However, the high costs of enzyme preparations significantly limit their applications in fermentation. In this study, the solid-state fermentation of millet bran was performed using a cellulase preparation and compound probiotics producing cellulase (CPPC), respectively. The results showed that both factors effectively destroyed the fiber structure, reduced the crude fiber content by 23.78% and 28.32%, respectively, and significantly increased the contents of beneficial metabolites and microorganisms. Moreover, CPPC could more effectively reduce the anti-nutrient factors and increase the content of anti-inflammatory metabolites. The correlation analysis revealed that Lactiplantibacillus and Issatchenkia had synergistic growth during fermentation. Overall, these results suggested that CPPC could replace cellulase preparation and improve antioxidant properties while reducing anti-nutrient factors of millet bran, thus providing a theoretical reference for the efficient utilization of agricultural by-products.

Deep soil microbial carbon metabolic function is important but often neglected: a study on the Songnen Plain reed wetland, Northeast China.

Soil microbial carbon metabolism is critical in wetland soil carbon cycling, and is also a research hotspot at present. However, most studies focus on the surface soil layer in the wetlands and the microorganisms associated with this layer. In this study, 0-75 cm soil profiles were collected from five widely separated reed wetlands in the Songnen Plain, which has a large number of middle-high latitude inland saline-sodic wetlands. The Biolog-ECO method was used to determine the carbon metabolic activity and functional diversity of soil microorganisms. The results showed that soil carbon metabolic activity decreased with increasing soil depth. The carbon metabolic activity of soil microorganisms in the 60-75 cm layer was approximately 57.41%-74.60% of that in the 0-15 cm layer. The soil microbial Shannon index and utilization rate of amines decreased with an increase in soil depth, while the Evenness index and utilization rate of polymers tended to increase with soil depth. Dissolved organic carbon (DOC) is the most important factor affecting microbial carbon source utilization preference, because microorganisms mainly obtain the carbon source from DOC. The result of the correlation analysis showed that the soil microbial carbon metabolic activity, Shannon index, and Evenness index significantly correlated with soil total carbon (TC), microbial biomass carbon (MBC), DOC, total nitrogen (TN), ammonium nitrogen (NH4 +-N), nitrate nitrogen (NO3 --N) contents, and electrical conductivity (EC). This study emphasized the important role of microbial carbon metabolic function in deep soil.

Single-Molecule Investigation of the Protein-Aptamer Interactions and Sensing Application Inside the Single Glass Nanopore.

Solid-state nanopores offer a nanoconfined space for a single-molecule sensing strategy. Evaluating the behavior of proteins and protein-related interactions at the single-molecule level is becoming more and more important for a better understanding of biological processes and diseases. In this work, the aptamer-functionalized nanopore was prepared as the sensing platform for kinetic analysis of the carcinoembryonic antigen (CEA) with its aptamers, which is an important cancer biomarker. CEA molecules were captured by the aptamers immobilized on the inner surface of the nanopore, and there was a complicated interaction between the CEA molecules and the aptamer, which is the process of association and dissociation. This could be used to measure the dynamics of aptamer-protein interactions without labeling. The kinetic analysis could be evaluated at the single-molecule level to interpret the dissociation constants of the binding and dissociation processes. Results showed that the translocation of CEA molecules in a functionalized nanopore had a deep blockades degree and long duration compared with nanopore modified with bare gold, which could be used for CEA sensing. This protein and protein-related interaction we designed provides new insights for evaluating the binding affinity, which will be beneficial for protein sensing and immunoassays.

Synergistic Effect of Oxoammonium Salt and Its Counterions for Fabricating Organic Electrochemical Transistors with Low Power Consumption.

The state-of-the-art poly(3,4-ethylenedioxythiophene):poly(styrene sulfonate) (PEDOT:PSS)-based organic electrochemical transistors (OECTs) are gaining importance for a variety of biological applications due to their mixed electronic and ionic conductivities featuring ion-to-electron conversion. A low operation voltage without sacrificing device performance is desired to realize long-term monitoring of biological activities. In the present work, oxoammonium salts with two different counterions (TEMPO+X-, where TEMPO = 2,2,6,6-tetramethylpiperidine-1-oxoammonium; X = Br- and TFSI-) are employed as secondary dopants to modulate the device performance. Both oxoammonium salts feature a distinct dopant concentration-dependent doping effect, allowing precise control in improving the performance of OECTs. A zero-gate bias, corresponding to the maximum transconductance, and a low threshold voltage are realized by optimizing the dopant concentrations. In addition, TEMPO+TFSI- dopant exerts great capability in modulating the work function and in morphology reconstruction of PEDOT:PSS, ensuring a well-matched work function at the gold electrode-channel material interface and condensed microstructure stacking with an edge-on orientation in the doped PEDOT:PSS films. The synergistic effect of TEMPO and the TFSI- counterion endows the device with superior performance to its counterparts due to the resultant higher µC* figure, benefiting from the efficient injection/extraction of holes at the interface and enhanced intra- and inter-chain carrier transport. The excellent device performance makes the OECT a promising neuromorphic device to mimic basic brain functions.

Dopant-Free Bithiophene-Imide-Based Polymeric Hole-Transporting Materials for Efficient and Stable Perovskite Solar Cells.

The development of hole-transport materials (HTMs) with high mobility, long-term stability, and comprehensive passivation is significant for simultaneously improving the efficiency and stability of perovskite solar cells (PVSCs). Herein, two donor-acceptor (D-A) conjugated polymers PBTI and PFBTI with alternating benzodithiophene (BDT) and bithiophene imide (BTI) units are successfully developed with desirable hole mobilities due to the good planarity and extended conjugation of molecular backbone. Both copolymers can be employed as HTMs with suitable energy levels and efficient defect passivation. Shortening the alkyl chain of the BTI unit and introducing fluorine atoms on the BDT moiety effectively enhances hole mobility and hydrophobicity of the HTMs, leading to improved efficiency and stability of PVSCs. As a result, the organic-inorganic hybrid PVSCs with PFBTI as the HTM deliver a power conversion efficiency (PCE) of 23.1% with enhanced long-term operational and ambient stability, which is one of the best efficiencies reported for PVSCs with dopant-free polymeric HTMs to date. Moreover, PFBTI can be applied in inorganic PVSCs and perovskite/organic tandem solar cells, achieving a high PCE of 17.4% and 22.2%, respectively. These results illustrate the great potential of PFBTI as an efficient and widely applicable HTM for cost-effective and stable PVSCs.

A solution-processed n-type conducting polymer with ultrahigh conductivity.

Conducting polymers (CPs) with high conductivity and solution processability have made great advances since the pioneering work on doped polyacetylene1-3, thus creating the new field of 'organic synthetic metals,4. Various high-performance CPs have been realized, which enable the applications of several organic electronic devices5,6. Nevertheless, most CPs exhibit hole-dominant (p-type) transport behaviour7,8, whereas the development of n-type analogues lags far behind and only a few exhibit metallic state, typically limited by low doping efficiency and ambient instability. Here we present a facilely synthesized highly conductive n-type polymer poly(benzodifurandione) (PBFDO). The reaction combines oxidative polymerization and in situ reductive n-doping, greatly increasing the doping efficiency, and a doping level of almost 0.9 charges per repeating unit can be achieved. The resultant polymer exhibits a breakthrough conductivity of more than 2,000 S cm-1 with excellent stability and an unexpected solution processability without extra side chains or surfactants. Furthermore, detailed investigations on PBFDO show coherent charge-transport properties and existence of metallic state. The benchmark performances in electrochemical transistors and thermoelectric generators are further demonstrated, thus paving the way for application of the n-type CPs in organic electronics.

Stochastic Collision Electrochemistry from Single Pt Nanoparticles: Electrocatalytic Amplification and MicroRNA Sensing.

Single-particle collisions have made many achievements in basic research, but challenges still exist due to their low collision frequency and selectivity in complex samples. In this work, we developed an "on-off-on" strategy based on Pt nanoparticles (PtNPs) that catalyze N2H4 collision signals on the surface of carbon ultramicroelectrodes and established a new method for the detection of miRNA21 with high selectivity and sensitivity. PtNPs catalyze the reduction of N2H4 on the surface of carbon ultramicroelectrodes to generate a stepped collision signal, which is in the "on" state. The single-stranded DNA paired with miRNA21 is coupled with PtNPs to form the complex DNA/PtNPs. Because PtNPs are covered by DNA, the electrocatalytic collision of N2H4 oxidation is inhibited. At this time, the signal is in the "off" state. When miRNA21 is added, the strong complementary pairing between miRNA21 and DNA destroys the electrostatic adsorption of DNA/PtNP conjugates and restores the electrocatalytic performance of PtNPs, and the signal is in the "on" state again. Based on this, a new method for detecting miRNA21 was established. It provides a new way for small-molecule sensing and has a wide range of applications in electroanalysis, electrocatalysis, and biosensing.

Anion-Doped Thickness-Insensitive Electron Transport Layer for Efficient Organic Solar Cells.

In organic solar cells, interfacial materials play essential roles in charge extraction, transportation, and collection. Currently, highly efficient and thickness-insensitive interfacial materials are urgently needed in printable large area module devices. Herein, water/alcohol-soluble conjugated polyelectrolyte PFNBT-Br, with medium bandgap based on benzothiadiazole, are doped by two alkali metal sodium salts, NaH2 PO2 , Na2 C2 O4 with different counter anions, to pursue high efficiency and thickness-insensitive electron-transport layers. Results show that the doping of electron-transport material can effectively promote the performance of the devices. Moreover, electron-transport layers doped by these salts with different counter anions show different behaviors in performances. Among which, the salt with oxalate anion C2 O4 2- (also named Ox2- ) shows much better device performance than the salt with hypophosphite anion (H2 PO2 - ), especially under the thick film condition (e.g., 50 nm). The greatly enhanced performances of interfacial material doped by Ox2- are due to reduced series resistance between the active layer material and the electrode, reduced dark-current, improved charge transport, and extraction efficiency, and decreased charge recombination for the devices at thick-film condition. These results demonstrated that n-doping could be a great potential strategy for making thickness-insensitive interfacial layers, besides, the performances can be further improved by carefully selecting salts.

Distinct Performance of Methylated SEPT9 in Upper and Lower Gastrointestinal Cancers and Combined Detection with Protein Markers.

Background: The performance of methylated SEPT9 (mSEPT9) in lower gastrointestinal (GI) cancer (colorectal cancer) has been extensively investigated; however, its performance in upper GI cancer (esophageal cancer and gastric cancer) and the comparison with lower GI cancer have rarely been studied. Methods: A total of 1854 subjects, including 344 upper GI cancer patients, 459 lower GI cancer patients, and 1051 noncancer subjects, were recruited in this prospective cohort study. A modified single polymerase chain reaction test for detecting mSEPT9 was used for plasma detection. Results: The sensitivity of mSEPT9 for upper and lower GI cancers was 45.3% and 74.8%, and the corresponding specificities were 85.6% and 86.5%, with areas under curve (AUC) of 0.71 and 0.80, respectively. mSEPT9 exhibited lower sensitivity in stage I than stage II-IV cancer, while no difference in sensitivity was observed for different locations in upper or lower GI cancer. No difference in sensitivity was found among gross classifications, pathological classifications, and differentiation in upper GI cancer, but a higher sensitivity in infiltrative cancer and moderate and poorly differentiated cancers was observed in the lower GI. No difference in sensitivity was found between male and female in both cancers, while sensitivity increased with age for both cancers. Cancer antigen 724 (CA724) showed the highest sensitivity for upper GI cancers, and carcinoembryonic antigen (CEA) showed the highest sensitivity for lower GI cancers. The combination of CA724 with mSEPT9 increased the sensitivity to 67.5% in upper GI cancers, and the combination of mSEPT9 with CEA increased the sensitivity to 85.4% in lower GI cancers, with an AUC of 0.90 and 0.95, respectively. Conclusions: mSEPT9 exhibited a higher sensitivity in lower GI cancers than upper GI cancers. The combination of mSEPT9 with protein markers significantly enhanced the detection sensitivity in both cancers.

Nanopore-Based Single-Entity Electrochemistry for the Label-Free Monitoring of Single-Molecule Glycoprotein-Boronate Affinity Interaction and Its Sensing Application.

Nanopipettes provide a promising confined space that enables advances in single-molecule analysis, and their unique conical tubular structure is also suitable for single-cell analysis. In this work, functionalized-nanopore-based single-entity electrochemistry (SEE) analysis tools were developed for the label-free monitoring of single-molecule glycoprotein-boronate affinity interaction for the first time, and immunoglobulin G (IgG, one of the important biomarkers for many diseases such as COVID-19 and cancers) was employed as the model glycoprotein. The principle of this method is based on a single glycoprotein molecule passing through 4-mercaptophenylboronic acid (4-MPBA)-modified nanopipettes under a bias voltage and in the meantime interacting with the boronate group from modified 4-MPBA. This translocation and affinity interaction process can generate distinguishable current blockade signals. Based on the statistical analysis of these signals, the equilibrium association constant (κa) of single-molecule glycoprotein-boronate affinity interaction was obtained. The results show that the κa of IgG in the confined nanopore at the single-molecule level is much larger than that measured in the open system at the ensemble level, which is possibly due to the enhanced multivalent synergistic binding in the restricted space. Moreover, the functionalized-nanopore-based SEE analysis tools were further applied for the label-free detection of IgG, and the results indicate that our method has potential application value for the detection of glycoproteins in real samples, which also paves way for the single-cell analysis of glycoproteins.