zhpsЖурнал прикладной спектроскопииZhurnal Prikladnoii Spektroskopii0514-7506B. I. Stepanov Institute of Physics of the National Academy of Scienceszhps-788Research ArticleСтатьиПЛАЗМОННЫЙ СЕНСОР ДЛЯ ОБНАРУЖЕНИЯ β-ЛАКТАМНЫХ АНТИБИОТИКОВ НА ОСНОВЕ КОНЪЮГИРОВАННОГО АНТИТЕЛА С НАНОЧАСТИЦАМИ ЗОЛОТАPLASMONIC SENSOR FOR DETECTION OF β-LACTAM ANTIBIOTICS BASED ON THE CONJUGATED ANTIBODY WITH GOLD NANOPARTICLESAghamirzaeiM.AghamirzaeiM.

Тебриз

Tabriz

KhiabaniM. S.KhiabaniM. S.

Тебриз

Tabriz

m.sowti@tabrizu.ac.irHamishehkarH.HamishehkarH.

Тебриз

Tabriz

MokarramR. R.MokarramR. R.

Тебриз

Tabriz

AmjadiM.AmjadiM.

Тебриз

Tabriz

Университет ТебризаUniversity of TabrizЦентр прикладных исследований медикаментов Тебризского медицинского университетаDrug Applied Research Center at Tabriz University of Medical Sciences202129012021881174(1)-174(10)Copyright © Aghamirzaei M., Khiabani M.S., Hamishehkar H., Mokarram R.R., Amjadi M., 20212021Aghamirzaei M., Khiabani M.S., Hamishehkar H., Mokarram R.R., Amjadi M.Aghamirzaei M., Khiabani M.S., Hamishehkar H., Mokarram R.R., Amjadi M.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://zhps.ejournal.by/jour/article/view/788

Предложен сенсор для обнаружения β-лактамных антибиотиков, основанный на использовании антител, связанных с наночастицами золота (НЧ-Au). Для колориметрического определения β-лактамных антибиотиков (ампициллина, амоксициллина, пенициллина G, оксациллина и карбенициллина) в качестве восстановителя задействованы НЧ-Au, синтезированные из экстракта листьев китайского салата. Методы рентгеновской дифракции, ИК-Фурье-спектроскопии, просвечивающей электронной микроскопии и динамического рассеяния света использованы для обнаружения кристалличности, идентификации функциональных групп, участвующих в синтезе НЧ-Au, и измерения их размера. В качестве наилучших значений выбраны pH 8, концентрация антител в 1 мл раствора НЧ-Au 8.4 мкг. С помощью спектрофотометра зарегистрированы спектры коллоидных НЧ-Au, антител, связанных с НЧ-Au, и антибиотиков (с концентрацией от 1 нМ до 1 мМ) в присутствии системы антитела–НЧ-Au. Показано, что с увеличением концентрации β-лактамных антибиотиков линия поглощения НЧ-Au сдвигается в красную область. При увеличении концентрации ампициллина, пенициллина G и карбенициллина длина волны максимума изменяется, однако по достижении некоторой концентрации антибиотиков (уровня насыщения) кривая выходит на плато, что указывает на влияние антител при обнаружении этих антибиотиков. Для амоксициллина концентрация насыщения намного выше, что указывает на особую роль антител при его обнаружении. Для оксациллина насыщение наступает очень быстро. Это демонстрирует, что антитела обладают чрезвычайно низкой способностью обнаружения данного антибиотика. Наличие антител позволяет обнаружить пять β-лактамных антибиотиков с чувствительностью до 1 нМ.

This study aims to detect β-lactam antibiotics using a conjugated antibody with gold nanoparticles (GNPs). For this purpose, the gold nanoparticles synthesized from Chinese lettuce leaf extract (as reductant) were used for the colorimetric detection of β-lactam antibiotics (such as ampicillin, amoxicillin, penicillin G, oxacillin, and carbenicillin). XRD, FTIR spectroscopy, TEM, and dynamic light scattering were utilized to detect the crystallinity, to identify functional groups involved in the synthesis of GNPs, and to measure the size of the GNPs; pH 8 and a concentration of 8.4 μg of antibody at 1 mL GNPs solution were selected as the best pH and concentration of antibody for the conjugation of antibody with GNPs. The maximum wavelengths of the colloidal GNPs, conjugation of antibody with GNPs, and detection of antibiotics (from 1 nM to 1 mM) with GNPs-PAb were recorded using a micro-volume spectrophotometer system. The results indicated that the localized surface plasmon resonance spectrometer absorption wavelength of GNPs red-shifted with increasing concentration of β-lactam antibiotics. With increasing concentration of ampicillin, penicillin G, and carbenicillin, the wavelength of maximum changed, and after saturation of antibiotics concentration, the curve reaches a plateau. This indicated that the antibody showed similar behavior in the detection of these antibiotics. But regarding amoxicillin, the saturation concentration is much higher, indicating that the antibody was more specific for its detection. In contrast, for oxacillin, saturation occurred very soon, which demonstrated that the antibody had an extremely low detection capability for this antibiotic. Finally, the results showed that the antibody was sensitive to 1 nM of the five β-lactam antibiotics studied.

спектрометр локализованного поверхностного плазмонного резонансаβ-лактамные антибиотикиполиклональные антитела к β-лактамунаночастицы золотаконъюгацияlocalized surface plasmon resonance spectrometerβ-lactam antibioticspolyclonal antibody of β-lactamgold nanoparticlesconjugationThis article is a part of a Ph.D. thesis approved at the University of Tabriz. This work was financially supported by Iran Nanotechnology Initiative Council (Grant code: 136472). The authors would like to thank Prof. Mohammad Javad Rasaee from Tarbiat Modares University, Dr. Seyed Mohammad Amini from Iran University of Medical Sciences, and Mr. Gholamreza Behrouzi, Mr. Babak Farshid, Miss Zahra Abolghasemi-Fakhri, and Mrs. Samira Tizchang for their very useful comments. We thank Rojan Azma Co. and its personnel for supporting research equipment. Finally, we thank Daneshmand Research & Development Institute and Mostazafan Foundation of Islamic Revolution.ReferencesV. Tamošiūnas, A. 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The authors declare that there are no conflicts of interest present.