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Approaches to Controlled Co-Amplification of Genes for Production of Biopharmaceuticals Study of the Insertion and Amplification Dynamics of Genetic Cassettes in the Genome of Chinese Hamster Ovary Cells during Co-Expression of Compatible Pair of Plasmidsстатья
Информация о цитировании статьи получена из
Web of Science,
Scopus
Статья опубликована в журнале из списка Web of Science и/или Scopus
Дата последнего поиска статьи во внешних источниках: 19 февраля 2020 г.
- Авторы: Kovnir S.V., Orlova N.A., Khodak Y.A., Kondrashova M.P., Gabibov A.G., Skryabin K.G., Vorobiev A.I., Vorobiev I.I.
- Журнал: Bulletin of Experimental Biology and Medicine
- Том: 163
- Номер: 2
- Год издания: 2017
- Издательство: Springer Nature
- Местоположение издательства: Switzerland
- Первая страница: 245
- Последняя страница: 249
- DOI: 10.1007/s10517-017-3776-0
- Аннотация: Plasmid vector family p1.1 based on non-coding regions of Chinese hamster housekeeping gene EEF1A and concatemer of Epstein-Barr virus terminal repeat increases the frequency of genome integration and provides rapid amplification of the target genes in the genome. For a pair of fluorescent proteins eGFP and mCherry it was shown that p1.1 vectors bearing dihydrofolate reductase and glutamine synthetase selection markers upon co-transfection into CHO DG44 cell line allow obtaining a polyclonal cell population in which 70% of cells express both genes. The subsequent one-step gene amplification of the genome-integrated genetic cassettes under the selective pressure of increased concentrations of methotrexate can increase the expression of both integrated genes up to 8.2% eGFP and 9.9% mCherry of total protein. This approach can be used for the development of cell lines for the production of functional heterodimeric proteins, e.g. polypeptide hormones and therapeutic antibodies.
- Добавил в систему: Орлова Надежда Александровна