Exploring mechanism of enzyme catalysis by on-chip transient kinetics coupled with global data analysis and molecular modeling

Hess, David; Dockalova, Veronika; Kokkonen, Piia; Bednář, David; Damborský, Jiřı́; deMello, Andrew J.; Prokop, Zbyněk; Stavrakis, Stavros

doi:10.1016/j.chempr.2021.02.011

Cited by 30 publications

(36 citation statements)

References 51 publications

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“…The microfluidic channel network used in the current study was designed and optimized to measure kinetic data on a millisecond to second time scale. Indeed, we have recently used a similar design for the study of enzyme kinetics on sub-millisecond time scales . Briefly, the channel structure allows for the rapid acceleration of droplets immediately after droplet formation to decrease mixing times.…”

Section: Resultsmentioning

confidence: 99%

“…Indeed, we have recently used a similar design for the study of enzyme kinetics on submillisecond time scales. 8 Briefly, the channel structure allows for the rapid acceleration of droplets immediately after droplet formation to decrease mixing times. This acceleration phase is followed by droplet deceleration to allow for efficient interrogation of droplets in the measurement channel.…”

Section: Kinetics Of Calcium Carbonate Precipitation In Dropletsmentioning

confidence: 99%

“…For example, in the case of reactive systems, it is possible to match volumetric flow rates and the locations where the system is interrogated in order to probe reaction times on a millisecond or sub-millisecond level. 8 Indeed, it should be realized that droplet-based microfluidic systems are best exploited when combined with sensitive and high-throughput detection methods that are able to probe individual droplet payloads. To this end, a diversity of detection methods have been successfully used with droplet-based microfluidic platforms to allow for the real-time characterization of encapsulated species and reactions.…”

Section: ■ Introductionmentioning

confidence: 99%

“…Indeed, the ability to create and homogenize temperature or solute gradients within confined droplets makes them uniquely suited for the study of fast reactions. , Moreover, mapping the position of each droplet as it moves downstream allows for the extraction of high-temporal resolution kinetic data, regardless of the acquisition time of the detection method being used. For example, in the case of reactive systems, it is possible to match volumetric flow rates and the locations where the system is interrogated in order to probe reaction times on a millisecond or sub-millisecond level . Indeed, it should be realized that droplet-based microfluidic systems are best exploited when combined with sensitive and high-throughput detection methods that are able to probe individual droplet payloads.…”

Section: Introductionmentioning

confidence: 99%

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In Situ X-ray Absorption Spectroscopy and Droplet-Based Microfluidics: An Analysis of Calcium Carbonate Precipitation

et al. 2021

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Droplet-based microfluidic systems are ideally suited for the investigation of nucleation and crystallization processes. To best leverage the features of such platforms (including exquisite time resolution and high-throughput operation), sensitive and in situ detection schemes are needed to extract real-time chemical information about all species of interest. In this regard, the extension of conventional (UV, visible, and infrared) optical detection schemes to the X-ray region of the electromagnetic spectrum is of high current interest, as techniques such as X-ray absorption spectroscopy (XAS) provide for the element-specific investigation of the local chemical environment. Accordingly, herein, we report for the first time the integration of millisecond droplet-based microfluidics with XAS. Such a platform allows for the sensitive acquisition of X-ray absorption data from picoliter-volume droplets moving at high linear velocities. Significantly, the high-temporal resolution of the droplet-based microfluidic platform enables unprecedented access to the early stages of the reaction. Using such an approach, we demonstrate in situ monitoring of calcium carbonate precipitation by extracting XAS spectra at the early time points of the reaction with a dead time as low as 10 ms. We obtain insights into the kinetics of the formation of amorphous calcium carbonate (ACC) as a first species during the crystallization process by monitoring the proportion of calcium ions converted into ACC. Within the confined and homogeneous environment of picoliter-volume droplets, the ACC content reaches 60% over the first 130 ms. More generally, the presented method offers new opportunities for the real-time monitoring of fast chemical and biological processes.

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Section: Resultsmentioning

confidence: 99%

Section: Kinetics Of Calcium Carbonate Precipitation In Dropletsmentioning

confidence: 99%

Section: ■ Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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In Situ X-ray Absorption Spectroscopy and Droplet-Based Microfluidics: An Analysis of Calcium Carbonate Precipitation

et al. 2021

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“…Bogdan Mateescu (University of Zürich) is leading the Purification of exRNA by Immuno-capture and Sorting using Microfluidics (PRISM) project together with Robert Raffai (University of California), Kendall Van Keuren-Jensen (TGEN), and Andrew deMello (ETH Zürich). This transdisciplinary team with expertise in lipoproteins, RNA and EVs biology ( Li et al., 2018a , 2018b ; Mateescu et al., 2017 ), exRNA sequencing ( Yeri et al., 2017 ), and microfluidics ( Hess et al., 2021 ; Holzner et al., 2021 ) aims to improve the sensitivity and specificity of exRNA-based diagnostic assays. First, the team will develop viscoelastic microfluidics platforms allowing the size-based separation of sub-micron particles to promote facile enrichment of exRNA nanocarriers from unprocessed biofluids.…”

Section: Introductionmentioning

confidence: 99%

Phase 2 of extracellular RNA communication consortium charts next-generation approaches for extracellular RNA research

Mateescu

Jones

Alexander³

et al. 2022

iScience

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Light‐Responsive Materials in Droplet Manipulation for Biochemical Applications

Cheng,

Kuan,

Lou

et al. 2024

Advanced Materials

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Miniaturized droplets, characterized by well‐controlled microenvironments and capability for parallel processing, have significantly advanced the studies on enzymatic evolution, molecular diagnostics, and single‐cell analysis. However, manipulation of small‐sized droplets, including moving, merging, and trapping of the targeted droplets for complex biochemical assays and subsequent analysis, is not trivial and remains technically demanding. Among various techniques, light‐driven methods stand out as a promising candidate for droplet manipulation in a facile and flexible manner, given the features of contactless interaction, high spatiotemporal resolution, and biocompatibility. This review therefore compiles an in‐depth discussion of the governing mechanisms underpinning light‐driven droplet manipulation. Besides, light‐responsive materials, representing the core of light‐matter interaction and the key character converting light into different forms of energy, are particularly assessed in this review. Recent advancements in light‐responsive materials and the most notable applications are comprehensively archived and evaluated. Continuous innovations and rational engineering of light‐responsive materials are expected to propel the development of light‐driven droplet manipulation, equip droplets with enhanced functionality, and broaden the applications of droplets for biochemical studies and routine biochemical investigations.This article is protected by copyright. All rights reserved

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Exploring mechanism of enzyme catalysis by on-chip transient kinetics coupled with global data analysis and molecular modeling

Cited by 30 publications

References 51 publications

In Situ X-ray Absorption Spectroscopy and Droplet-Based Microfluidics: An Analysis of Calcium Carbonate Precipitation

In Situ X-ray Absorption Spectroscopy and Droplet-Based Microfluidics: An Analysis of Calcium Carbonate Precipitation

Phase 2 of extracellular RNA communication consortium charts next-generation approaches for extracellular RNA research

Light‐Responsive Materials in Droplet Manipulation for Biochemical Applications

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