transmedТрансляционная медицинаTranslational Medicine2311-44952410-5155Almazov National Medical Research Centre, Saint Petersburg, Russia10.18705/2311-4495-2025-12-1-67-79TJMXSBtransmed-971Research ArticleБИОИНЖЕНЕРНЫЕ И БИОИНФОРМАТИЧЕСКИЕ ТЕХНОЛОГИИBIOENGINEERING AND BIOINFORMATICSАффинное выделение экзосом с помощью магнитных наночастицSpecific isolation of exosomes using magnetic nanosphereМоисееваЕ. О.MoiseevaE. O.

Моисеева Екатерина Олеговна, аспирант 

Москва 

Ekaterina O. Moiseeva, PhD student 

 Moscow 

КожевниковаД. Д.KozhevnikovaD. D.

Кожевникова Дарья Дмитриевна, аспирант 

Москва 

Daria D. Kozhevnikova, PhD student 

 Moscow 

СергеевИ. С.SergeevI. S.

Сергеев Игорь Сергеевич, аспирант 

Москва 

Igor S. Sergeev, PhD student 

Moscow 

БлиндманМ. А.BlindmanM. A.

Блиндман Мария Александровна, аспирант 

Москва 

Maria A. Blindman, PhD student 

Moscow 

ЗабороваО. В.ZaborovaO. V.

Заборова Ольга Владимировна, к.х.н., старший научный сотрудник, химический факультет 

Москва 

Olga V. Zaborova, PhD (Chem.), Senior Researcher, Faculty of Chemistry 

Moscow 

ЧернышевВ. С.ChernyshevV. S.

Чернышев Василий Сергеевич, к.т.н., заведующий лабораторией 

Москва 

Vasiliy S. Chernyshev, PhD (Tech.), Head of Laboratory 

Moscow 

ЯковлевА. А.YakovlevA. A.

Яковлев Александр Александрович, д.б.н., старший научный сотрудник 

Москва 

Alexander A. Yakovlev, Dr. Sci. (Biol.), Senior Researcher 

Moscow

ГоринД. А.GorinD. A.

Горин Дмитрий Александрович, д.х.н., профессор 

Москва 

Dmitry A. Gorin, Dr. Sci. (Chem.), Professor 

Moscow

https://orcid.org/0000-0002-8302-5646ЯщенокА. М.YashchenokA. M.

Ященок Алексей Михайлович, д.ф.-м.н., доцент 

Большой бул., д. 30, Москва, 121205 

Alexey M. Yashchenok, Dr. Sci. (Phys.-Math.), Associate Professor 

Bolshoy Boulevard, 30, p. 1, Moscow, 121205 

a.yashchenok@skoltech.ruАвтономная некоммерческая образовательная организация высшего образования «Сколковский институт науки и технологий»РоссияSkolkovo Institute of Science and Technology, Skolkovo Innovation CenterRussian FederationФедеральное государственное бюджетное образовательное учреждение высшего образования «Московский государственный университет имени М. В. Ломоносова»РоссияLomonosov Moscow State UniversityRussian FederationФедеральное государственное бюджетное учреждение «Национальный медицинский исследовательский центр акушерства, гинекологии и перинатологии имени академика В. И. Кулакова» Министерства здравоохранения Российской ФедерацииРоссияAcademician V. I. Kulakov National Medical Research Centre for Obstetrics, Gynecology and PerinatologyRussian FederationФедеральное государственное бюджетное учреждение науки «Институт высшей нервной деятельности и нейрофизиологии Российской академии наук»РоссияInstitute of Higher Nervous Activity and Neurophysiology of Russian Academy of SciencesRussian Federation202507052025121Внеклеточные везикулы: большие перспективы маленьких объектов6779Copyright © Моисеева Е.О., Кожевникова Д.Д., Сергеев И.С., Блиндман М.А., Заборова О.В., Чернышев В.С., Яковлев А.А., Горин Д.А., Ященок А.М., 20252025Моисеева Е.О., Кожевникова Д.Д., Сергеев И.С., Блиндман М.А., Заборова О.В., Чернышев В.С., Яковлев А.А., Горин Д.А., Ященок А.М.Moiseeva E.O., Kozhevnikova D.D., Sergeev I.S., Blindman M.A., Zaborova O.V., Chernyshev V.S., Yakovlev A.A., Gorin D.A., Yashchenok A.M.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://transmed.almazovcentre.ru/jour/article/view/971

Актуальность. Экзосомы относятся к классу внеклеточных везикул, которые секретируются клетками и циркулируют в биологических жидкостях, опосредуя межклеточную коммуникацию и передачу сигналов в различных патологических процессах, в том числе в модуляции развития раковых опухолей и опухолевого микроокружения. Клетки раковых опухолей продуцируют повышенное количество экзосом уже на начальных стадиях заболевания. Причем характерные для опухоли белковые маркеры могут встречаться в составе генерируемых экзосом, что делает последние перспективным объектом для детекции заболевания методом жидкостной биопсии. Основным препятствием для внедрения экзосом как маркеров развития заболеваний является отсутствие удобного, недорогого и быстрого метода для их выделения и детекции. Цель. Разработка платформы для выделения экзосом и детектирования белковых мембранных маркеров, изолированных экзосом. Материалы и методы. Разработаны магнитные наночастицы, функционализированные анти-EpCAM аптамером, для избирательного захвата субпопуляции экзосом опухолевых клеток. Эффективность магнитных наночастиц протестирована на экзосомах, полученных из EpCAM-позитивной линии клеток рака кишечника (HT29). Для обеспечения высокой чувствительности анализа экзосом из патологически измененных клеток, везикулы предварительно метили липофильным флуоресцентным красителем (3,3’диоктадецилоксакарбоцианин). Результаты. Показано, что магнитные наночастицы, связанные с аптамером анти-EpCAM, эффективно захватывали экзосомы из культуральной среды HT29. Методом проточной флуоресцентной цитометрии получен предел обнаружения порядка 107 экзосом/мл. Заключение. Настоящее исследование демонстрирует эффективный метод избирательного выделения экзосом от конкретного типа клеток, перспективный для современной диагностики и мониторинга онкологических заболеваний.

Background. Exosomes belong to the class of extracellular vesicles that are secreted by cells and circulate in biological fluids, mediating intercellular communication and signaling in various pathological processes, including modulation of cancer development and tumor microenvironment. Exosomes released by cancer cells at early stages of disease progression expressing specific membrane markers, which makes exosomes a promising biomarker of the disease by liquid biopsy. The main obstacle to the use of exosomes as markers of disease development is the lack of a convenient, inexpensive and rapid method for their isolation and detection. Objective. The aim of this study was to develop a platform for exosome isolation and detection of exosome protein membrane markers. Design and methods. In this work, magnetic nanoparticles functionalized with anti-EpCAM aptamer were developed for selective capture of tumor cell exosomes. The efficiency of magnetic nanoparticles was tested on exosomes derived from EpCAM-positive colon cancer cell line (HT29). To ensure high sensitivity of exosomes from pathologically cells, vesicles were pre-labeled with a lipophilic fluorescent dye (3,3’ dioctadecyloxacarbocyanine). Results. It was shown that magnetic nanoparticles bound with anti-EpCAM aptamer effectively captured exosomes from the HT29 culture medium. The detection limit of about 107 exosomes/ml was obtained using flow cytometry method. Conclusion. This study demonstrates an effective method for selective isolation of exosomes from cancer cell, which is promising for diagnostics and monitoring of oncological diseases.

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