Droplet microfluidics as a high-throughput approach for enzyme
kinetics studies

a 2023

Droplet microfluidics as a high-throughput approach for enzyme kinetics studies

SEDLÁK, Michal, Marta PELCOVÁ, Lenka KOHÚTOVÁ a Zdeněk GLATZ

Základní údaje

Originální název

Droplet microfluidics as a high-throughput approach for enzyme kinetics studies

Autoři

SEDLÁK, Michal, Marta PELCOVÁ, Lenka KOHÚTOVÁ a Zdeněk GLATZ

Vydání

Doctoral conference 2023, 2023

Další údaje

Typ výsledku

Konferenční abstrakt

Utajení

není předmětem státního či obchodního tajemství

Klíčová slova česky

Kapičková mikrofluidika; Enyzymová kinetika; Laboratoř na čipu

Klíčová slova anglicky

Droplet microfluidics; Enzyme kinetics; Lab on a chip

Změněno: 27. 10. 2023 11:28, Mgr. Michal Sedlák

Anotace

V originále

Microfluidic systems are recently undergoing rapid development as a high-throughput method useful in bioanalytical applications. Droplet microfluidics is emerging as a promising method in this field. Basic concept of droplet microfluidics is generation of large amount of water based droplets in immiscible oil phase. Since the droplets are isolated and protected by the oil and can be individually manipulated, they can be regarded as separate microreactors. Compared to the conventional technologies, microfluidics offers several advantages such as reduced reagent consumption, shorter analyses times, and rapid production of data in given time. The research of enzyme kinetics is of high interest as many enzymes play their role in human diseases and are potential therapeutical drug targets. The aim is to develop convenient droplet microfluidics based methods for performing enzyme assays with the mentioned advantages. The "hearth" of the device is typically polydimethylsiloxane (PDMS) chip, serving as droplet generator and incubation channel, connected to precise syringe pumps. The enzyme reaction can be monitored using synthetic fluorogenic substrates, offering high sensitivity. For this purpose the microscope is mounted with excitation light source, set of optical filters and photomultiplier tube serving as a detector. The setup allows to measure the reaction rate simply by focusing on different distances in the incubation channel, corresponding to different reaction times. First optimizations of uniform-sized droplets generation as well as pilot study with E. coli beta-galactosidase with fluorogenic substrate as a model system has been done. In the following three years of the doctoral studies the main efforts will be focused on further miniaturization, optimization of the method, and its application for clinically and pharmacologically interesting enzymes.

Návaznosti

CZ.02.2.69/0.0/0.0/16_018/0002605, interní kód MU

Název: Bioanalytické technologie ve výzkumu a klinické diagnostice (Akronym: DSP Bioanalytické technologie)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Bioanalytické technologie ve výzkumu a klinické diagnostice, PO 2 Rozvoj vysokých škol a lidských zdrojů pro výzkum a vývoj

Citovat

SEDLÁK, Michal, Marta PELCOVÁ, Lenka KOHÚTOVÁ a Zdeněk GLATZ. Droplet microfluidics as a high-throughput approach for enzyme kinetics studies. In Doctoral conference 2023. 2023.

@proceedings{2330457,
   author = {Sedlák, Michal and Pelcová, Marta and Kohútová, Lenka and Glatz, Zdeněk},
   booktitle = {Doctoral conference 2023},
   keywords = {Droplet microfluidics; Enzyme kinetics; Lab on a chip},
   title = {Droplet microfluidics as a high-throughput approach for enzyme kinetics studies},
   year = {2023}
}

TY  - CONF
ID  - 2330457
AU  - Sedlák, Michal - Pelcová, Marta - Kohútová, Lenka - Glatz, Zdeněk
PY  - 2023
TI  - Droplet microfluidics as a high-throughput approach for enzyme kinetics studies
KW  - Droplet microfluidics
KW  - Enzyme kinetics
KW  - Lab on a chip
N2  - Microfluidic systems are recently undergoing rapid development as a high-throughput method useful in bioanalytical applications. Droplet microfluidics is emerging as a promising method in this field. Basic concept of droplet microfluidics is generation of large amount of water based droplets in immiscible oil phase. Since the droplets are isolated and protected by the oil and can be individually manipulated, they can be regarded as separate microreactors. Compared to the conventional technologies, microfluidics offers several advantages such as reduced reagent consumption, shorter analyses times, and rapid production of data in given time. The research of enzyme kinetics is of high interest as many enzymes play their role in human diseases and are potential therapeutical drug targets. The aim is to develop convenient droplet microfluidics based methods for performing enzyme assays with the mentioned advantages. The "hearth" of the device is typically polydimethylsiloxane (PDMS) chip, serving as droplet generator and incubation channel, connected to precise syringe pumps. The enzyme reaction can be monitored using synthetic fluorogenic substrates, offering high sensitivity. For this purpose the microscope is mounted with excitation light source, set of optical filters and photomultiplier tube serving as a detector. The setup allows to measure the reaction rate simply by focusing on different distances in the incubation channel, corresponding to different reaction times. First optimizations of uniform-sized droplets generation as well as pilot study with E. coli beta-galactosidase with fluorogenic substrate as a model system has been done. In the following three years of the doctoral studies the main efforts will be focused on further miniaturization, optimization of the method, and its application for clinically and pharmacologically interesting enzymes.
ER  -

SEDLÁK, Michal, Marta PELCOVÁ, Lenka KOHÚTOVÁ a Zdeněk GLATZ. Droplet microfluidics as a high-throughput approach for enzyme kinetics studies. In \textit{Doctoral conference 2023}. 2023.

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