Исследование функциональной активности химически модифицированной рекомбинантной бутирилхолинэстеразы человека in vitro и in vivo

Выводы.

1. Разработана эффективная система экспрессии рекомбинантной БуХЭ человека в клетках линии СНО, позволяющая получать до 30 мг/л активного фермента в олигомерной форме.

2. Полученный рекомбинантный фермент обладает сравнимыми структурно-функциональными характеристиками с препаратом БуХЭ плазмы крови человека.

3. Впервые успешно осуществлено химическое полисиалирование гидролитического фермента. Подобраны условия реакции, позволяющие получать конъюгат рчБуХЭ-СА027 с 80%-ным выходом. Показано, что данная модификация не изменяет кинетические характеристики фермента, что свидетельствует об интактности его активного центра.

4. Применение химического полисиалирования существенно улучшило фармакокинетические параметры рекомбинантного фермента, увеличив время полувыведения модифицированной рчБуХЭ в 5,5 раз.

5. Полисиалированный препарат показал высокую эффективность в качестве профилактического биологического антидота. Защитный индекс по веществу УЫ составил 4,2 ЛД50, что сопоставимо с БуХЭ плазмы крови человека.

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