Single Protein Molecule Detection by Glass Nanopores

Li, Wenhong; Bell, Nicholas A. W.; Hernández‐Ainsa, Silvia; Thacker, Vivek V.; Thackray, Alana M.; Bujdoso, Raymond; Keyser, Ulrich F.

doi:10.1021/nn4004567

Cited by 241 publications

(267 citation statements)

References 35 publications

Supporting

Mentioning

260

Contrasting

Order By: Relevance

“…This range is slightly larger than the protein size range used in a similar setup using glass nanocapillaries published by Li et al, while our measurements were done at a constant Bessel filter frequency of 10 kHz. 36 The ability to shrink glass nanopores has important advantages like tailoring the glass nanopore diameter to the size of the target molecule (ESI Fig. 2 †).…”

Section: Discussionmentioning

confidence: 99%

Probing the size of proteins with glass nanopores

et al. 2014

View full text Add to dashboard Cite

a Single molecule studies using nanopores have gained attention due to the ability to sense single molecules in aqueous solution without the need to label them. In this study, short DNA molecules and proteins were detected with glass nanopores, whose sensitivity was enhanced by electron reshaping which decreased the nanopore diameter and created geometries with a reduced sensing length. Further, proteins having molecular weights (MW) ranging from 12 kDa to 480 kDa were detected, which showed that their corresponding current peak amplitude changes according to their MW. In the case of the 12 kDa ComEA protein, its DNA-binding properties to an 800 bp long DNA molecule was investigated. Moreover, the influence of the pH on the charge of the protein was demonstrated by showing a change in the translocation direction. This work emphasizes the wide spectrum of detectable molecules using nanopores from glass nanocapillaries, which stand out because of their inexpensive, lithography-free, and rapid manufacturing process.

show abstract

Section: Discussionmentioning

confidence: 99%

Probing the size of proteins with glass nanopores

et al. 2014

View full text Add to dashboard Cite

show abstract

“…Importantly, they can be rapidly and inexpensively laser pulled from glass capillaries, resulting in a sharp tip constituting a single nanopore, which can be used as single-molecule label-free sensors, [10][11][12][13][14] in the same way as conventional solid-state nanopores. Due to their high-aspect ratio geometry and exceptionally sharp tips, nanopipettes offer an important advantage over conventional nanopores as they can be easily adapted for use in single cell interrogation 15,16 and as a macroscopic delivery vehicle for intracellular injection.…”

Section: Introductionmentioning

confidence: 99%

“…In fact nanopipettes are predominantly used as an analytical platform for the detection of single molecules by transporting molecules from the outside reservoir to the inside of the nanopipette. 10,12,13,[24][25][26][27] . However, this does not take advantage of utilizing the nanopipette as a label-free single-molecule delivery vehicle.…”

Section: Introductionmentioning

confidence: 99%

On-Demand Delivery of Single DNA Molecules Using Nanopipets

et al. 2015

View full text Add to dashboard Cite

Understanding the behavioral properties of single molecules or larger scale populations interacting with single molecules is currently a hotly pursued topic in nanotechnology. This arises from the potential such techniques have in relation to applications such as targeted drug delivery, early stage detection of disease, and drug screening. Although label and label-free single molecule detection strategies have existed for a number of years, currently lacking are efficient methods for the controllable delivery of single molecules in aqueous environments. In this article we show both experimentally and from simulations that nanopipets in conjunction with asymmetric voltage pulses can be used for label-free detection and delivery of single molecules through the tip of a nanopipet with "on-demand" timing resolution. This was demonstrated by controllable delivery of 5 kbp and 10 kbp DNA molecules from solutions with concentrations as low as 3 pM.

show abstract

“…Nanopore quantification can outperform the traditional surface assays in sensitivity because it does not rely on reversible hybridization. The most common nanopore types are short and small (<1 nm) "soft" protein pores, [127][128][129] conic solid-state pores, 130,131 and a hybrid of the two as in the Oxford Nanopore technology with a gating motor protein at the end of a solid-state nanopore.…”

Section: Polymer Nanopores and Nanopipettesmentioning

confidence: 99%

“…[147][148][149][150][151][152] For example, we can pack 25 nm to 100 nm oligo-functionalized Au, silica and polystyrene NPs into iontrack etched polyethylene terephthalate (PET) polymer conic nanopores, 134,135 and into conic silica nanocapillaries pulled by laser assisted drawing. 130,136,137 We have studied the NP assembly mechanism with two-dimensional nano-wedge models fabricated by focused ion beam milling. 153 In a conic nanopore, NPs larger than the conic tip diameter ($100 nm) can be assembled by electrophoresis either on its tip or through its base.…”

Section: Polymer Nanopores and Nanopipettesmentioning

confidence: 99%

Future microfluidic and nanofluidic modular platforms for nucleic acid liquid biopsy in precision medicine

et al. 2016

View full text Add to dashboard Cite

Nucleic acid biomarkers have enormous potential in non-invasive diagnostics and disease management. In medical research and in the near future in the clinics, there is a great demand for accurate miRNA, mRNA, and ctDNA identification and profiling. They may lead to screening of early stage cancer that is not detectable by tissue biopsy or imaging. Moreover, because their cost is low and they are non-invasive, they can become a regular screening test during annual checkups or allow a dynamic treatment program that adjusts its drug and dosage frequently. We briefly review a few existing viral and endogenous RNA assays that have been approved by the Federal Drug Administration. These tests are based on the main nucleic acid detection technologies, namely, quantitative reverse transcription polymerase chain reaction (PCR), microarrays, and next-generation sequencing. Several of the challenges that these three technologies still face regarding the quantitative measurement of a panel of nucleic acids are outlined. Finally, we review a cluster of microfluidic technologies from our group with potential for point-of-care nucleic acid quantification without nucleic acid amplification, designed to overcome specific limitations of current technologies. We suggest that integration of these technologies in a modular design can offer a low-cost, robust, and yet sensitive/selective platform for a variety of precision medicine applications.

show abstract

Single Protein Molecule Detection by Glass Nanopores

Cited by 241 publications

References 35 publications

Probing the size of proteins with glass nanopores

Probing the size of proteins with glass nanopores

On-Demand Delivery of Single DNA Molecules Using Nanopipets

Future microfluidic and nanofluidic modular platforms for nucleic acid liquid biopsy in precision medicine

Contact Info

Product

Resources

About