A novel method to purify adenovirus based on increasing salt concentrations in buffer

Ma, Jinhu; Su, Chao; Wang, Xilei; Shu, Yongheng; Hu, Shichuan; Zhao, Chengzhi; Kuang, Yueting; Chen, Yanwei; Li, Yuhua; Wei, Yuquan; Prof, Ping Cheng

doi:10.1016/j.ejps.2019.105090

Cited by 8 publications

(6 citation statements)

References 28 publications

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“…In the present study, during the upstream process for large-scale preparation of adenovirus, Triton X-100 was added (with a final concentration of 0.25%) to the bioreactor for lysing the cells to release the adenovirus. 19 In this method, host cells can be lysed in a closed environment of the bioreactor, leading to reduced probability of contamination of the adenovirus feed, followed by linear scale-up. The cell lysate harvested at the same time can be well connected with the downstream adenovirus purification process.…”

Section: Discussionmentioning

confidence: 99%

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

et al. 2020

Molecular Therapy - Methods & Clinical Development

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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.

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Section: Discussionmentioning

confidence: 99%

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

et al. 2020

Molecular Therapy - Methods & Clinical Development

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“…However, ultracentrifugation is not convenient for high-scale production and good manufacturing practices (GMP) adaptation. A combination of chromatographic methods based on capture antibodies, ionic exchange, size exclusion, hydrophobic interaction, and immobilized metal affinity columns has been described [43,44]. Methods that contribute to the enrichment in full vector particles are especially indicated for HC-AdV [45,46].…”

Section: Purificationmentioning

confidence: 99%

High-Capacity Adenoviral Vectors: Expanding the Scope of Gene Therapy

Ricobaraza

González-Aparicio

Mora-Jimenez

et al. 2020

IJMS

101

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The adaptation of adenoviruses as gene delivery tools has resulted in the development of high-capacity adenoviral vectors (HC-AdVs), also known, helper-dependent or “gutless”. Compared with earlier generations (E1/E3-deleted vectors), HC-AdVs retain relevant features such as genetic stability, remarkable efficacy of in vivo transduction, and production at high titers. More importantly, the lack of viral coding sequences in the genomes of HC-AdVs extends the cloning capacity up to 37 Kb, and allows long-term episomal persistence of transgenes in non-dividing cells. These properties open a wide repertoire of therapeutic opportunities in the fields of gene supplementation and gene correction, which have been explored at the preclinical level over the past two decades. During this time, production methods have been optimized to obtain the yield, purity, and reliability required for clinical implementation. Better understanding of inflammatory responses and the implementation of methods to control them have increased the safety of these vectors. We will review the most significant achievements that are turning an interesting research tool into a sound vector platform, which could contribute to overcome current limitations in the gene therapy field.

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“…65 One study also suggests that when UF/DF is performed with increasing salt concentrations, clearance of serum proteins and HCPs is significant, thus increasing loading amount into a consequent chromatographic column and improving binding of VLPs to packing media. 66 Continuous diafiltration, consisting of multistage arrangement of membranes, can potentially be used to self-assemble VLPs more efficiently. In this approach, output from the preceding stage is used as input for the following stage, while diluent is added either in a co-current or counter-current manner.…”

Section: Continuous Filtration Technologiesmentioning

confidence: 99%

Current status and future challenges in transitioning to continuous bioprocessing of virus‐like particles

Mittal

Banerjee

Lua

et al. 2021

J of Chemical Tech & Biotech

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Virus-like particles (VLPs) are bioengineered protein complexes that can serve as prophylactic vaccines due to favorable immunological characteristics like size, repetitive surface geometry and ability to induce both innate and adaptive immune responses, without being infectious. Large-scale manufacturing of VLPs presents some interesting and unique challenges associated with removal of refractory contaminants to attain high product purity, improvement in structural homogeneity and colloidal stability. Recent advancements in production and purification of VLP-based vaccines, such as application of monolithic multicolumn chromatography and inline concentration and diafiltration, have set a stage for implementation of continuous bioprocessing for commercial manufacturing of VLPs. The benefits of continuous bioprocessing have been well demonstrated for biotherapeutics, particularly monoclonal antibodies and small peptides. With the growing demand for affordable vaccines, the biopharmaceutical industry has shown increasing interest in establishing end-to-end continuous platforms. This article reviews major developments that have occurred in continuous processing for the manufacturing of VLPs. The major problems inherent in transitions from batch to continuous processing are also discussed, along with potential strategies to overcome the challenges.

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A novel method to purify adenovirus based on increasing salt concentrations in buffer

Cited by 8 publications

References 28 publications

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

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

High-Capacity Adenoviral Vectors: Expanding the Scope of Gene Therapy

Current status and future challenges in transitioning to continuous bioprocessing of virus‐like particles

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