Ma Jinhu, Su Chao, Hu Shichuan, Chen Yanwei, Shu Yongheng, Yue Dan, Zhang Bin, Qi Zhongbing, Li Suli, Wang Xilei, Kuang Yueting, Cheng Ping
State Key Laboratory of Biotherapy and Cancer Center/Collaborative Innovation Center for Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, PR China.
Mol Ther Methods Clin Dev. 2020 Aug 20;19:35-46. doi: 10.1016/j.omtm.2020.08.013. eCollection 2020 Dec 11.
To ensure the high purity and biological activity of the adenovirus vector to be used for clinical applications, a stable and linearly scalable preparation method is highly imperative. During the adenovirus-harvesting process, the Triton X-100-based lysis method possesses the advantages of higher efficiency as well as easier linearization and amplification. Most Triton X-100 can be removed from the adenovirus sample by chromatographic purification. However, there is no report that a small amount of residual Triton X-100, present in adenovirus sample, can affect the particle integrity, infectivity, and structure of adenoviruses. Here, we found that although residual Triton X-100 affected the short-term stability, purity, infectivity, and structure of adenoviruses at 37°C, it did not hamper these properties of adenoviruses at 4°C. This study suggests that although the Triton X-100-based lysis method is a simple, efficient, and easy-to-scale process for lysing host cells to release the adenovirus, the storage conditions of adenovirus products must be taken into consideration.
为确保用于临床应用的腺病毒载体具有高纯度和生物活性,一种稳定且可线性扩展的制备方法至关重要。在腺病毒收获过程中,基于 Triton X - 100 的裂解方法具有效率更高以及线性化和扩增更容易的优点。大多数 Triton X - 100 可通过色谱纯化从腺病毒样品中去除。然而,尚无报道表明腺病毒样品中存在的少量残留 Triton X - 100 会影响腺病毒的颗粒完整性、感染性和结构。在此,我们发现尽管残留的 Triton X - 100 在 37°C 时会影响腺病毒的短期稳定性、纯度、感染性和结构,但在 4°C 时它不会妨碍腺病毒的这些特性。本研究表明,尽管基于 Triton X - 100 的裂解方法是一种用于裂解宿主细胞以释放腺病毒的简单、高效且易于扩展的过程,但必须考虑腺病毒产品的储存条件。