Optimization on cell lysis and capture process of human adenovirus type 5 produced in suspension HEK293 cells

IF 1.4 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS

Protein expression and purification Pub Date : 2025-02-04 DOI:10.1016/j.pep.2025.106686

Sha Yi, Xiaoxu Gu, Youping Jin, Fei Wang, Junjun Jiang

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引用次数: 0

Abstract

Adenovirus (Adv) is increasingly recognized for its significance in the fields of gene therapy and viral vector vaccines. The diverse applications in clinical trials and fundamental research necessitate the development of environmentally and economically sustainable purification processes that are straightforward and scalable for both academic and industrial contexts. In the initial segment of this study, we evaluated the lysis efficiency of polysorbate 20 (PS20) in comparison to polysorbate 80 (PS80). Our findings indicated that the viability HEK293 could be reduced to approximately 13 %, with a detectable Adv5 concentration of average 1.62 × 10⁹ ifu/mL in the supernatant after an incubation in 1.0 % (w/w) PS20 or PS80 buffer. In the subsequent portion of this research, we employed a high-throughput static binding capacity (SBC) screening tool in conjunction with on-column dynamic binding capacity (DBC) validation to concurrently assess the binding capacity of Adv5 on nine different types of anion exchange media. The results demonstrated that both Sartobind Q membrane and POROS 50HQ resin exhibited binding capacities exceeding 1.0 × 10¹³ vp/mL under the testing conditions.

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来源期刊

Protein expression and purification 生物-生化研究方法

CiteScore

3.70

自引率

6.20%

发文量

120

审稿时长

32 days

期刊介绍： Protein Expression and Purification is an international journal providing a forum for the dissemination of new information on protein expression, extraction, purification, characterization, and/or applications using conventional biochemical and/or modern molecular biological approaches and methods, which are of broad interest to the field. The journal does not typically publish repetitive examples of protein expression and purification involving standard, well-established, methods. However, exceptions might include studies on important and/or difficult to express and/or purify proteins and/or studies that include extensive protein characterization, which provide new, previously unpublished information.