antibioticsАнтибиотики и ХимиотерапияAntibiot Khimioter = Antibiotics and Chemotherapy0235-2990ООО «Издательство ОКИ»antibiotics-683Research ArticleОРИГИНАЛЬНЫЕ СТАТЬИORIGINAL PAPERSФлуоресцентный поляризационный иммуноанализ для экспрессного контроля содержания антибиотиков: разработка и характеристика на примере хлорамфениколаFluorescence Polarization Immunoassay for Express Control of Antibiotic Levels: Design and Characteristics for Chloramphenicol, as an ExampleЕреминС. А.EreminS. A.noemail@neicon.ruХанО. Ю.KhanO. Yu.noemail@neicon.ruПисаревВ. В.PisarevV. V.noemail@neicon.ruЗвереваЕ. А.ZverevaE. A.noemail@neicon.ruЖердевА. В.ZherdevA. V.noemail@neicon.ruДзантиевБ. Б.DzantievB. B.noemail@neicon.ruМосковский государственный университет им. М. В. ЛомоносоваРоссияM. V. Lomonosov Moscow State UniversityRussian FederationНаучно-производственный центр «Пробиотек»РоссияScientific-Production Center «Probiotek»Russian FederationИнститут биохимии им. А. Н. Баха, Федеральный исследовательский центр «Фундаментальные основы биотехнологии» РАНРоссияA. N. Bakh Institute of Biochemistry, Federal Research Centre «Fundamentals of Biotechnology» Russian Academy of SciencesRussian Federation201613052020619-102227Copyright © ООО «Издательство ОКИ», 20202020ООО «Издательство ОКИ»ООО «Издательство ОКИ»https://www.antibiotics-chemotherapy.ru/jour/about/submissions#copyrightNoticehttps://www.antibiotics-chemotherapy.ru/jour/article/view/683

Рассмотрены особенности флуоресцентного поляризационного иммуноанализа (ФПИА) как средства экспрессного контроля наличия и содержания антибиотиков в различных видах проб, его преимущества по сравнению с другими аналитическими методами. На примере детекции антибиотика хлорамфеникола рассмотрены этапы разработки аналитической методики, определения её параметров. Анализ основан на конкурентном взаимодействии антител против хлорамфеникола с конъюгатом хлорамфеникол-флуорофор и потенциально содержащимся в тестируемой пробе свободным антибиотиком. Представлены экспериментальные результаты сравнения методик ПФИА хлорамфеникола, реализованных с использованием двух конъюгатов, которые отличаются длиной мостикового участка между функциональными группами антибиотика и флуорофора (флуоресцеина). Охарактеризованы требования к выбору концентраций антител и конъюгата, обеспечивающих высокочувствительную детекцию. Предел обнаружения хлорамфеникола при использовании разработанного ФПИА - 10 нг/мл, диапазон определяемых концентраций - от 20 нг/мл до 10 мкг/мл. Время определения - 10 мин.

Characteristics of the fluorescence polarization immunoassay (FPIA) as a mean for express control of antibiotic levels in various specimens and its advantages vs. other analytical tests are described. The developmental stages of the analytical procedure and its parameters are considered for chlorampnenicol as an example. The analysis is based on competitive interaction of anti-chloramphenicol antibodies with the chloramphenicol-fluorophore conjugate and the potential free chloramphenicol in the specimen. The experimental results of the comparison of the chloramphenicol FPIA with the use of two conjugates differing in the length of the bridge length between the antibiotic functional groups and fluorophore (fluorescein) are presented. The requirements to the choice of the antibody and conjugate concentrations providing highly sensitive detection are characterized. The detection limit of chloramphenicol in the FPIA was 10 ng/ml and the determination of the concentrations ranged from 20 ng/ml to 10 mcg/ml. The time of the assay was 10 min.

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The authors declare that there are no conflicts of interest present.