Nature Biotechnology, Год журнала: 2025, Номер unknown
Опубликована: Март 17, 2025
Язык: Английский
Journal of Proteome Research, Год журнала: 2024, Номер 23(6), С. 1871 - 1882
Опубликована: Май 7, 2024
The coevolution of liquid chromatography (LC) with mass spectrometry (MS) has shaped contemporary proteomics. LC hyphenated to MS now enables quantification more than 10,000 proteins in a single injection, number that likely represents most specific human cells or tissues. Separations by ion mobility (IMS) have recently emerged complement and further improve the depth Given theoretical advantages speed robustness IMS comparison LC, we envision ongoing improvements paired may eventually make obsolete, especially when combined targeted simplified analyses, such as rapid clinical proteomics analysis defined biomarker panels. In this perspective, describe need for faster might drive transition, current state direct infusion proteomics, discuss some technical challenges must be overcome fully complete transition entirely gas phase
Язык: Английский
Процитировано
6
Sensors and Actuators Reports, Год журнала: 2024, Номер 8, С. 100225 - 100225
Опубликована: Июль 17, 2024
Nanopores play essential roles in biological processes, such as ion channels and pumps cellular membranes, technological applications DNA sequencing. Advancements nanofabrication techniques have enabled the routine integration of nanopores into solid-state devices, resulting a plethora analytical applications. This review explores recent developments nanopore-enabled electrochemical systems, which transcended traditional resistive pulse sensing to offer novel capabilities single-entity studies, stimulus-responsive gating, point-of-care diagnostics. We highlight studies on design utility nanopore electrode arrays, serve nanocontainers capable isolating analyzing single entities, extend discussion hierarchically organized, systems that regulate species transport across nanopores, enriching analytes for ultrasensitive detection. In addition, we utilization probe-assisted sensing, demonstrating its efficacy selectively binding detecting target molecules ions. Finally, outline future directions nanopore-based enhance robustness, achieve high-throughput analysis, incorporate artificial intelligence materials data promising transformative impacts diagnostics research.
Язык: Английский
Процитировано
6
ACS Nano, Год журнала: 2024, Номер 18(36), С. 25155 - 25169
Опубликована: Авг. 27, 2024
Steviol glycosides (SGs) are a class of high-potency noncalorie natural sweeteners made up common diterpenoid core and varying glycans. Thus, the diversity glycans in composition, linkage, isomerism results tremendous structural complexity SG family, which poses challenges for precise identification leads to fact that SGs frequently used mixtures their variances biological activity remain largely unexplored. Here we show wild-type aerolysin nanopore can detect discriminate diverse species through modulable electro-osmotic flow effect at varied applied voltages. At low voltages, neutral molecule was drawn stuck pore entrance due an energy barrier around R220 sites. The ensuing binding events enable majority species. Increasing voltage break cause translocation events, allowing unambiguous several pairs differing by only one hydroxyl group recognition accumulation from multiple sensing regions Based on data 15 SGs, deep learning-based artificial intelligence (AI) model created process individual blockage achieving rapid, automated, single-molecule quantification real samples. This work highlights value analysis complex glycans-containing glycosides, as well potential sensitive rapid quality assurance glycoside products with use AI.
Язык: Английский
Процитировано
5
Expert Review of Proteomics, Год журнала: 2025, Номер unknown
Опубликована: Март 19, 2025
Liquid chromatography-tandem mass spectrometry (LC-MS/MS) has been a leading method for proteomics thirty years. Advantages provided by LC-MS/MS are offset significant disadvantages, including cost. Recently, several non-mass spectrometric methods have emerged, but little information is available about their capacity to analyze the complex mixtures routine spectrometry. We review recent non-mass-spectrometric sequencing proteins and peptides, using nanopores, degradation, reverse translation, short-epitope mapping, with comments on bioinformatics challenges, fundamental limitations, areas where new technologies will be more or less competitive LC-MS/MS. In addition conventional literature searches, instrument vendor websites, patents, webinars, preprints were also consulted give up-to-date picture. Many promising. However, demonstrations that they outperform in terms of peptides identified not yet published, astute observers note important especially relating dynamic range single-molecule measurements mixtures. Still, even if performance emerging proves inferior LC-MS/MS, low cost could create different kind revolution: dramatic increase number biology laboratories engaging forms research.
Язык: Английский
Процитировано
0
Analytica Chimica Acta, Год журнала: 2025, Номер unknown, С. 343955 - 343955
Опубликована: Март 1, 2025
Язык: Английский
Процитировано
0
Advanced Science, Год журнала: 2024, Номер 11(28)
Опубликована: Апрель 19, 2024
In recent decades, there has been a significant increase in the application of single-molecule electrical analysis platforms studying proteins and peptides. These advanced methods have potential for deep investigation enzymatic working mechanisms accurate monitoring dynamic changes protein configurations, which are often challenging to achieve ensemble measurements. this work, prominent research progress peptide protein-related studies surveyed using electronic devices with single-molecule/single-event sensitivity, including junctions, field-effect transistors, nanopores. particular, successful commercial nanopores DNA sequencing made it one most promising techniques at level. From single peptides complexes, correlation between their characteristics, structures, biological functions is gradually being established. This enables distinguish different molecular configurations these biomacromolecules through real-time life activities, significantly improving understanding underlying various processes.
Язык: Английский
Процитировано
4
Proteomes, Год журнала: 2024, Номер 12(2), С. 14 - 14
Опубликована: Апрель 19, 2024
With growing recognition and acknowledgement of the genuine complexity proteomes, we are finally entering post-proteogenomic era. Routine assessment proteomes as inferred correlates gene sequences (i.e., canonical ‘proteins’) cannot provide necessary critical analysis systems-level biology that is needed to understand underlying molecular mechanisms pathways or identify most selective biomarkers therapeutic targets. These requirements demand at level proteoforms/protein species, actual active players. Currently, only highly refined integrated integrative top-down proteomics (iTDP) enables analytical depth routine, comprehensive, quantitative proteome assessments across widest range proteoforms inherent native systems. Here a broad perspective field, taking in historical current realities, establish more balanced understanding where field has come from (in particular during ten years since Proteomes was launched), issues, how things likely need proceed if deep analyses succeed. We base this our firm belief best proteomic reflect, closely possible, sample moment sampling. also seek emphasise future approaches based on exploitation complementarity currently successful approaches. This emphasises continuously evaluate further optimize established approaches, avoid complacency thinking expectations but promote careful development introduction new notably those address proteoforms. Above all, wish rigorous focus quality must override largely values speed; latter would certainly be nice, could thus effectively, routinely, quantitatively assessed. Alas, composed proteoforms, not species can amplified directly mirror genes ‘canonical’). The problem hard, accept it such, payoff playing longer game promise far biomarkers, drug targets, truly personalised even individualised medicine.
Язык: Английский
Процитировано
4
Nature Methods, Год журнала: 2024, Номер unknown
Опубликована: Ноя. 18, 2024
Язык: Английский
Процитировано
4
Journal of the American Chemical Society, Год журнала: 2025, Номер unknown
Опубликована: Янв. 2, 2025
Nanopores are promising sensors for glycan analysis with the accurate identification of complex glycans laying foundation nanopore-based sequencing. However, their applicability toward continuous sequencing has not yet been demonstrated. Here, we present a proof-of-concept by combining nanopore technology glycosidase-hydrolyzing reactions. By continuously monitoring changes in characteristic current generated translocation hydrolysis products through nanopore, sequence can be accurately identified based on specificity glycosidases. With machine learning, improved accuracy to over 98%, allowing reliable determination consecutive building blocks and glycosidic linkages chains while reducing need operator expertise. This approach was validated real samples, calibrated using hydrophilic interaction chromatography-high-performance liquid chromatography (HILIC-HPLC) mass spectrometry (MS). We achieved ten units natural chains, which provided first evidence feasibility nanopore-glycosidase-compatible system Compared traditional methods, this strategy enhances efficiency 5-fold. Additionally, introduced concept 'inverse sequencing', focuses electrical signal rather than monosaccharide identification. eliminates reliance fingerprint libraries typically required putative 'forward hydrolysis' strategies. When challenges both inverse strategies' addressed, will pave way establishing at single-molecule level.
Язык: Английский
Процитировано
0
Analytical Chemistry, Год журнала: 2025, Номер unknown
Опубликована: Янв. 22, 2025
Nanopore sensing is widely used for single-molecule detection, originally applied to nucleic acids and now extended protein sensing. Our study focuses on the complex conformational changes of peptides in nanopores, which may have implications peptide fingerprinting identification. Specifically, we investigated interaction a β-hairpin (SV28) within an α-hemolysin (αHL) nanopore. experiments revealed that SV28 captured via dielectrophoresis exhibits long dwell times nanopore, leading multiple current blockade levels. Unlike DNA hairpins, showed non-sequential transitions among four distinct This behavior indicates dynamics nanopores cannot be simply modeled along single reaction coordinate. findings provide insights into peptide-nanopore interactions, are potentially useful developing nanopore-based identification technologies.
Язык: Английский
Процитировано
0