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A simplified ATP method for the rapid control of cell viability in a freeze-dried BCG vaccineстатья
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: 4 декабря 2016 г.
- Авторы: Ugarova Natalia N., Lomakina Galina Yu, Modestova Yulia, Chernikov Sergey V., Vinokurova Natalia V., Оtrashevskaya Elena V., Gorbachev Vyacheslav Y.
- Журнал: Journal of Microbiological Methods
- Том: 130
- Год издания: 2016
- Издательство: Elsevier BV
- Местоположение издательства: Netherlands
- Первая страница: 48
- Последняя страница: 53
- DOI: 10.1016/j.mimet.2016.08.027
- Аннотация: We propose a simple and cost-effective ATP method for controlling the specific activity of a freeze-dried BCG vaccine. A freeze-dried BCG vaccine is reconstituted with 1 ml saline and incubated for 15 min at room temperature and then for 1 h at 37 °C. The vaccine is then treated with apyrase to remove extracellular ATP. After that, the cells are lysed with DMSO and the ATP content in the lysate is measured by the bioluminescence method. To implement the method, we developed a kit that requires no time-consuming preparation before the analysis. We demonstrated the linear relationship between the experimental values of the specific activity (106 CFU/mg) and intracellular ATP content (ATP, pmol/mg) for different batches of the studied BCG vaccines; the proportionality coefficient was К = 0.36 ± 0.02. We proposed a formula for calculating the specific activity from the measured content of intracellular ATP (ATP, pmol/mg). The comparison of the measured and calculated values of the specific activity (106 CFU/mg) shows that these values are similar; their differences fall within the allowable range of deviations for the specific activity values of the BCG vaccine.
- Добавил в систему: Модестова Юлия Александровна