Сибирский федеральный университет

Тип публикации: статья из журнала

Год издания: 2017

Идентификатор DOI: 10.1134/S0003683817070043

Ключевые слова: bioluminescence, luciferase, microfluidics, microfluidic chip, enzymatic bioassay

Аннотация: A number of approaches have been proposed and tested to transfer enzymatic reactions into the functional elements of microfluidic chips on the example of the bienzyme bioluminescent reaction involving NAD(P)H:FMN-oxidoreductase and luciferase. Measurement of the catalytic activity of these enzymes (under the influence of pollutants) is the basis of enzymatic bioassay of various liquids. It was found that all of the components of the reaction must be placed in the same cell of the chip to improve the reproducibility of the measurements. The use of starch gel as a carrier for immobilization and gelatin as a scaffold in the reactor of the chip enables the preservation of enzyme activity in the course of sealing the chip at room temperature. It is shown that the components of the reaction should be vigorously stirred in a microfluidic chip reactor to improve the efficiency of the analysis. As a result of the studies, a prototype of microfluidic chip based on the enzymatic bioluminescent reaction is proposed. It is characterized by a detection limit of copper sulfate of 3 mu M that corresponds to the sensitivity of traditional lux-biosensors based on living cells. The analysis time is reduced to 1 min, and the analysis can be performed by individuals without special laboratory skills.

Журнал: APPLIED BIOCHEMISTRY AND MICROBIOLOGY

Выпуск журнала: Vol. 53, Is. 7

Номера страниц: 775-780

ISSN журнала: 00036838

Место издания: NEW YORK

Издатель: MAIK NAUKA/INTERPERIODICA/SPRINGER

• Lukyanenko K.A. (Siberian Fed Univ, Krasnoyarsk 660041, Russia)
• Denisov I.A. (Siberian Fed Univ, Krasnoyarsk 660041, Russia)
• Yakimov A.S. (Siberian Fed Univ, Krasnoyarsk 660041, Russia)
• Esimbekova E.N. (Siberian Fed Univ, Krasnoyarsk 660041, Russia; Russian Acad Sci, Inst Biophys, Siberian Branch, Krasnoyarsk 660036, Russia)
• Belousov K.I. (St Petersburg Inst Fine Mech & Opt, St Petersburg 197101, Russia)
• Bukatin A.S. (Siberian Fed Univ, Krasnoyarsk 660041, Russia)
• Kukhtevich I.V. (Siberian Fed Univ, Krasnoyarsk 660041, Russia)
• Sorokin V.V. (Siberian Fed Univ, Krasnoyarsk 660041, Russia)
• Evstrapov A.A. (St Petersburg Inst Fine Mech & Opt, St Petersburg 197101, Russia; Inst Analyt Instrumentat, St Petersburg 198095, Russia)
• Belobrov P.I. (Siberian Fed Univ, Krasnoyarsk 660041, Russia; Russian Acad Sci, Inst Biophys, Siberian Branch, Krasnoyarsk 660036, Russia)

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