Open Access

Biosensors, volume 13, issue 1, pages 47

Immobilization of Firefly Bioluminescent System: Development and Application of Reagents

Esimbekova E. N. ^{1, 2}

Kirillova Maria A ¹

Kratasyuk Valentina A. ^{1, 2}

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Institute of Fundamental Biology and Biotechnology, Siberian Federal University, 79 Svobodny Avenue, 660041 Krasnoyarsk, Russia |

Institute of Biophysics SB RAS, 50/50 Akademgorodok, 660036 Krasnoyarsk, Russia |

Publication type: Journal Article

Publication date: 2022-12-28

Multidisciplinary Digital Publishing Institute (MDPI)

Journal: Biosensors

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Impact factor: 5.4

ISSN: 20796374, 0265928X

DOI: 10.3390/bios13010047

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General Medicine

Clinical Biochemistry

Analytical Chemistry

Biotechnology

Instrumentation

Biomedical Engineering

Engineering (miscellaneous)

Abstract

The present study describes the method of preparing reagents containing firefly luciferase (FLuc) and its substrate, D-luciferin, immobilized into gelatin gel separately or together. The addition of stabilizers dithiothreitol (DTT) and bovine serum albumin (BSA) to the reagent is a factor in achieving higher activity of reagents and their stability during storage. The use of immobilized reagents substantially simplifies the procedure of assay for microbial contamination. The mechanism of action of the reagents is based on the relationship between the intensity of the bioluminescent signal and the level of ATP contained in the solution of the lysed bacterial cells. The highest sensitivity to ATP is achieved by using immobilized FLuc or reagents containing separately immobilized FLuc and D-luciferase. The limit of detection of ATP by the developed reagents is 0.3 pM, which corresponds to 20,000 cells·mL−1. The linear response range is between 0.3 pM and 3 nM ATP. The multicomponent reagent, containing co-immobilized FLuc and D-luciferin, shows insignificantly lower sensitivity to ATP—0.6 pM. Moreover, the proposed method of producing an immobilized firefly luciferin-luciferase system holds considerable promise for the development of bioluminescent biosensors intended for the analysis of microbial contamination.

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Citations by journals

	1
International Journal of Molecular Sciences	International Journal of Molecular Sciences, 1, 50% International Journal of Molecular Sciences 1 publication, 50%
Micromachines	Micromachines, 1, 50% Micromachines 1 publication, 50%
	1

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	1 2
Multidisciplinary Digital Publishing Institute (MDPI)	Multidisciplinary Digital Publishing Institute (MDPI), 2, 100% Multidisciplinary Digital Publishing Institute (MDPI) 2 publications, 100%
	1 2

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Esimbekova E. N. et al. Immobilization of Firefly Bioluminescent System: Development and Application of Reagents // Biosensors. 2022. Vol. 13. No. 1. p. 47.

GOST all authors (up to 50) Copy

Esimbekova E. N., Kirillova M. A., Kratasyuk V. A. Immobilization of Firefly Bioluminescent System: Development and Application of Reagents // Biosensors. 2022. Vol. 13. No. 1. p. 47.

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RIS Copy

TY - JOUR

DO - 10.3390/bios13010047

UR - https://doi.org/10.3390%2Fbios13010047

TI - Immobilization of Firefly Bioluminescent System: Development and Application of Reagents

T2 - Biosensors

AU - Esimbekova, E. N.

AU - Kirillova, Maria A

AU - Kratasyuk, Valentina A.

PY - 2022

DA - 2022/12/28 00:00:00

PB - Multidisciplinary Digital Publishing Institute (MDPI)

SP - 47

IS - 1

VL - 13

SN - 2079-6374

SN - 0265-928X

ER -

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BibTex Copy

@article{2022_Esimbekova,

author = {E. N. Esimbekova and Maria A Kirillova and Valentina A. Kratasyuk},

title = {Immobilization of Firefly Bioluminescent System: Development and Application of Reagents},

journal = {Biosensors},

year = {2022},

volume = {13},

publisher = {Multidisciplinary Digital Publishing Institute (MDPI)},

month = {dec},

url = {https://doi.org/10.3390%2Fbios13010047},

number = {1},

pages = {47},

doi = {10.3390/bios13010047}

}

MLA

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Esimbekova, E. N., et al. “Immobilization of Firefly Bioluminescent System: Development and Application of Reagents.” Biosensors, vol. 13, no. 1, Dec. 2022, p. 47. https://doi.org/10.3390%2Fbios13010047.

Found error?

Publisher

Multidisciplinary Digital Publishing Institute (MDPI)

Journal

Biosensors

Quartile SCImago

Quartile WOS

Impact factor

5.4

ISSN

20796374 (Electronic)
0265928X (Print)

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