Single antibody–antigen interactions monitored via transient ionic current recording using nanopore sensors

Ying, Yi‐Lun; Yu, Ru‐Jia; Hu, Yong‐Xu; Gao, Rui

doi:10.1039/c7cc03927a

Cited by 55 publications

(49 citation statements)

References 31 publications

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“…Transient PPIs can be measured using a resistive-pulse technique 8 and solid-state nanopores that are large enough to permit tethering of protein receptors on their internal surface. 9, 10 However, accurate identification of the anchoring site for tethered protein receptors remains difficult, which limits the sensitivity of single-molecule measurements. An alternative approach to these shortcomings is the detection of reversible PPIs using an engineered protein nanopore.…”

mentioning

confidence: 99%

Real-time measurement of protein–protein interactions at single-molecule resolution using a biological nanopore

2018

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Editors summary Single-molecule analyses of transient protein-protein interactions are enabled using an engineered biological nanopore.Protein-protein interactions (PPIs) are essential for many cellular processes. However, the transient nature of these PPIs is difficult to quantitate due to numerous limitations of available methods. We engineered a genetically-encoded sensor for real-time sampling of transient PPIs at single-molecule resolution. Our sensor comprises a truncated outer membrane protein pore, a flexible tether, a protein receptor and a peptide adapter. When a protein ligand present in solution binds to the receptor reversible capture and release events of the receptor can be measured as current transitions between two open substates of the pore. Importantly, the binding and release of the receptor by a protein ligand can be unambiguously discriminated in a complex sample containing fetal bovine serum. Our selective nanopore sensor could be applied for single-molecule protein detection, could form the basis for a nanoproteomics platform or might be adapted to build tools for protein profiling and biomarker discovery.

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mentioning

confidence: 99%

Real-time measurement of protein–protein interactions at single-molecule resolution using a biological nanopore

2018

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“…Approaches to slow down the translocation speed have been successfully reported in nanopore sensing of proteins at the single-molecule level. For instance, the functionalisation of quartz nanopores with specific antibodies was performed to monitor the interaction between single α-foetal proteins and its antibody at single-molecule level, after the affinity binding of both molecules in the nanopore [229]. A controllable gate voltage in a nanopore extended field-effect transistor allowed the slowing down of the transport of molecules through the nanopore.…”

Section: Nanoporesmentioning

confidence: 99%

Advanced Nanoscale Approaches to Single-(Bio)entity Sensing and Imaging

Neves

Martín‐Yerga

2018

Biosensors

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Individual (bio)chemical entities could show a very heterogeneous behaviour under the same conditions that could be relevant in many biological processes of significance in the life sciences. Conventional detection approaches are only able to detect the average response of an ensemble of entities and assume that all entities are identical. From this perspective, important information about the heterogeneities or rare (stochastic) events happening in individual entities would remain unseen. Some nanoscale tools present interesting physicochemical properties that enable the possibility to detect systems at the single-entity level, acquiring richer information than conventional methods. In this review, we introduce the foundations and the latest advances of several nanoscale approaches to sensing and imaging individual (bio)entities using nanoprobes, nanopores, nanoimpacts, nanoplasmonics and nanomachines. Several (bio)entities such as cells, proteins, nucleic acids, vesicles and viruses are specifically considered. These nanoscale approaches provide a wide and complete toolbox for the study of many biological systems at the single-entity level.

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“…In a typical setup, an external voltage drives a biomolecule through a nanometer-size pore in a thin supporting membrane, causing a characteristic temporary change in the translocation ionic current. Nanopore technology can be applied to assay the spatial structure [23][24][25], dynamic changes [25][26][27][28][29], and biochemical characteristics [30,31]. It is not necessary to carry out chemical modification, surface adsorption, and calibration object insertion, which may affect the activity of the sample.…”

Section: Introductionmentioning

confidence: 99%

Recognition of Bimolecular Logic Operation Pattern Based on a Solid-State Nanopore

Han

Zhang

Weng

et al. 2020

Sensors

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Nanopores have a unique advantage for detecting biomolecules in a label-free fashion, such as DNA that can be synthesized into specific structures to perform computations. This method has been considered for the detection of diseased molecules. Here, we propose a novel marker molecule detection method based on DNA logic gate by deciphering a variable DNA tetrahedron structure using a nanopore. We designed two types of probes containing a tetrahedron and a single-strand DNA tail which paired with different parts of the target molecule. In the presence of the target, the two probes formed a double tetrahedron structure. As translocation of the single and the double tetrahedron structures under bias voltage produced different blockage signals, the events could be assigned into four different operations, i.e., (0, 0), (0, 1), (1, 0), (1, 1), according to the predefined structure by logic gate. The pattern signal produced by the AND operation is obviously different from the signal of the other three operations. This pattern recognition method has been differentiated from simple detection methods based on DNA self-assembly and nanopore technologies.

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Single antibody–antigen interactions monitored via transient ionic current recording using nanopore sensors

Cited by 55 publications

References 31 publications

Real-time measurement of protein–protein interactions at single-molecule resolution using a biological nanopore

Real-time measurement of protein–protein interactions at single-molecule resolution using a biological nanopore

Advanced Nanoscale Approaches to Single-(Bio)entity Sensing and Imaging

Recognition of Bimolecular Logic Operation Pattern Based on a Solid-State Nanopore

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