J 2022

Droplet-Based Microfluidic Temperature-Jump Platform for the Rapid Assessment of Biomolecular Kinetics

YANG, Tianjin, Alessia VILLOIS, Antonín KUNKA, Fulvio GRIGOLATO, Paolo AROSIO et. al.

Základní údaje

Originální název

Droplet-Based Microfluidic Temperature-Jump Platform for the Rapid Assessment of Biomolecular Kinetics

Autoři

YANG, Tianjin, Alessia VILLOIS, Antonín KUNKA (203 Česká republika, domácí), Fulvio GRIGOLATO, Paolo AROSIO, Zbyněk PROKOP (203 Česká republika, garant, domácí), Andrew DEMELLO a Stavros STAVRAKIS

Vydání

Analytical chemistry, American Chemical Society, 2022, 0003-2700

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10400 1.4 Chemical sciences

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Odkazy

Impakt faktor

Impact factor: 7.400

Kód RIV

RIV/00216224:14310/22:00128417

Organizační jednotka

Přírodovědecká fakulta

UT WoS

000888138400001

Klíčová slova anglicky

Biotechnology; Fluid dynamics; Kinetics; Liquids; Nanofibers

Štítky

Příznaky

Mezinárodní význam, Recenzováno
Změněno: 31. 1. 2023 08:46, Mgr. Marie Šípková, DiS.

Anotace

V originále

Protein folding, unfolding, and aggregation are important in a variety of biological processes and intimately linked to “protein misfolding diseases”. The ability to perform experiments at different temperatures allows the extraction of important information regarding the kinetics and thermodynamics of such processes. Unfortunately, conventional stopped-flow methods are difficult to implement, generate limited information, and involve complex sample handling. To address this issue, we present a temperature-controlled droplet-based microfluidic platform that allows measurement of reaction kinetics on millisecond to second timescales and at temperatures between ambient and 90 °C. The utility of the microfluidic platform for measuring fast biomolecular kinetics at high temperatures is showcased through the investigation of the unfolding kinetics of haloalkane dehalogenases and the elongation of fibrils composed of the amyloid β peptide associated with Alzheimer’s disease. In addition, a deep-ultraviolet (UV) fluorescence microscope was developed for the on-chip recording of protein intrinsic fluorescence spectrum originating from aromatic amino acid residues. We envision that the developed optofluidic platform will find wide applicability in the analysis of biological processes, such as protein refolding and phase separation.

Návaznosti

EF17_043/0009632, projekt VaV
Název: CETOCOEN Excellence
857560, interní kód MU
(Kód CEP: EF17_043/0009632)
Název: CETOCOEN Excellence (Akronym: CETOCOEN Excellence)
Investor: Evropská unie, CETOCOEN Excellence, Spreading excellence and widening participation