Сборка трастузумаб (герцептин)-связывающих наночастиц вируса табачной мозаики в листьях: Nicotiana benthamiana тема диссертации и автореферата по ВАК РФ 03.01.06, кандидат биологических наук Петруня, Игорь Валерьевич

• Петруня, Игорь Валерьевич
• кандидат биологических наук
• 2010

Оглавление диссертации кандидат биологических наук Петруня, Игорь Валерьевич

1. ОГЛАВЛЕНИЕ.

2. СПИСОК СОКРАЩЕНИЙ.

4. ЦЕЛИ ИССЛЕДОВАНИЯ.

Продукция целевых белков в растениях с использованием транзиентной системы экспрессии.

Агроинфскция.

Транзиентная система экспрессии на основе АдгоЬаМег'шт Ште/аЫепх.

Векторы на основе вирусов растений.

Тобамовирусы.

Потивирусы.

Бромовирусы.

Комовирусы.

Геминивирусы.

Система экспрессии MagnICON.

Вирусоподобные частицы, получаемые в растениях.

Экспонирование эпитопов вакцинных белков на поверхности вирионов растительных вирусов.

5. МАТЕРИАЛЫ И МЕТОДЫ.

6. РЕЗУЛЬТАТЫ И ОБСУЖДЕНИЕ.

Исследование сборки трастузумаб-связывающих наночастиц ВТМ в листьях N. ЬешЬатгапа.

Изучение возможности трансформации клеток мыши при бактериемии А. Ште/аает.

7. ВЫВОДЫ.

8. БЛАГОДАРНОСТИ.

Заключение диссертации по теме «Биотехнология (в том числе бионанотехнологии)», Петруня, Игорь Валерьевич

7. ВЫВОДЫ

1. Разработанный метод получения трастузумаб-связывающих наночастиц рекомбинантного ВТМ (рВТМ) в листьях Шсойапа ЬеШкатгапа с использованем коагроинъекции экспрессионных векторов на базе ВТМ и ХВК обеспечивает высокий выход безопасного для окружающей среды рекомбинантного вируса.

2. Определены условия эффективной сборки рВТМ: выбранный пептид должен иметь физико-химические параметры нативного БО, не содержать остатков цистеина и предполагаемых сайтов гликозилирования.

3. Установлено, что наночастицы рВТМ с экспонированными на поверхности модифицированными пептидами НЕК2/пеи стабильны при хранении и обладают более высокой по сравнению с пептидами иммуногенностью.

4. Полученные наночастицы рВТМ могут быть использованы для аффинной очистки продуцируемого в растениях фитогерцептина.

5. Рекомбинантный ВТМ не индуцирует образование НЕ112/пеи-специфичных антител у мышей, что свидетельствует о ранее неизвестной множественности сайтов связывания трастузумаба с нативным НЕЯ2/пеи.

6. Технология получения рВТМ с различными экспонированным эпитопами может быть использована при поиске и селекции антигенных детерминант НЕК2/пеи для создания антираковой вакцины.

7. Показана биологическая безопасность А. Ште/аЫет для млекопитающих: при экспериментальной бактериемии А. Ште/аЫет не вызывает трансформацию клеток органов мыши.

8. БЛАГОДАРНОСТИ

Автор глубоко признателен своему научному руководителю Юрию Леонидовичу Дорохову за постоянное внимание, терпение и неизменную поддержку при выполнении этой работы. Также хотелось бы выразить отдельную благодарность Ольге Юрьевне Фроловой за неоценимый вклад в формировании автора как самостоятельного исследователя, а также бесценный опыт, переданный в течение всех лет работы в лаборатории. Выражаю благодарность Татьяне Валерьевне Комаровой за помощь в подготовке материалов для диссертации, участие в обсуждении результатов и ценные замечания, а также за помощь в освоении необходимых лабораторных методов. Хотелось бы поблагодарить Евгения Валерьевича Шеваля, за содействие в проведении электронномикроскопических исследований и обработку полученных данных, а также Вячеславу Станиславовичу Косорукову за оказанную помощь в постановке экспериментов. Автор признателен также профессору Юрию Юрьевичу Глеба (Icon Genetics AG) и профессору Виталию Цитовскому (Stony Brook University) за помощь в обсуждении результатов и ценные замечания. Большую благодарность автор выражает всем сотрудникам кафедры вирусологии, а именно Шевелевой Анне, Зверевой Анне Сергеевне, Шварцу Антону Марковичу, Иванову Петру Алексеевичу, Скурату Евгению Владимировичу, Тюлькиной Лидии Германовне за теплую, дружескую атмосферу, внимание, помощь и поддержку.

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