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The Effect of Residual Triton X-100 on Structural Stability and Infection Activity of Adenovirus Particles

Jinhu Ma, Chao Su, Shichuan Hu, Yanwei Chen, Yongheng Shu, Dan Yue, Bin Zhang, Zhongbing Qi, Suli Li, Xilei Wang, Yueting Kuang, Ping Cheng

2020Molecular Therapy — Methods & Clinical Development16 citationsDOIOpen Access PDF

Abstract

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. 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.

Topics & Concepts

Triton X-100LysisInfectivityAdenoviridaeChromatographyChemistryResidualBiologyVirologyMolecular biologyVirusPulmonary surfactantRecombinant DNAMathematicsBiochemistryAlgorithmGeneVirus-based gene therapy researchCAR-T cell therapy researchViral Infectious Diseases and Gene Expression in Insects