Solubilization and refolding of inclusion bodies by detergents.

IF 1.2 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS

Protein expression and purification Pub Date : 2025-12-01 Epub Date: 2025-08-07 DOI:10.1016/j.pep.2025.106791

Tsutomu Arakawa, Teruo Akuta, Daisuke Ejima, Kouhei Tsumoto

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引用次数: 0

Abstract

Recombinant proteins play many important roles in development of biological reagents and biopharmaceuticals. Here, we will mainly review refolding of recombinant proteins when expressed in inclusion bodies, although strategies to enhance soluble expression are described as an alternative to refolding inclusion bodies. These strategies include, but not limited to, adding chemical chaperones in cell culture media, modifying cell lysis buffer and using solubility-enhacing fusion tags. Another solubility enhancement was to generate lipid complex for membrane proteins that form insoluble proteins without lipid. Among various solubilization and refolding technologies, those using denaturant, alkaline pH and pressure are also desribed, while we focus on solubilization and refolding using detergents, which are effective and cost-friendly. Sodium dodecylsulfate, lauroyl-glutamate, sarkosyl and cetyltrimethylammonium have been extensively used, as summarized in this review. Slow or step-wise removal of denaturants or ionic detergents used to solubilize appears to play a critical role in successful refolding by maintaining the solubility of proteins during refolding. In alkaline refolding, slow pH adjustment also helps maintain protein solubility. In pressure refolding, small amount of guanidine hydrochloride assisted refolding.

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洗涤剂对包裹体的增溶和再折叠作用。

重组蛋白在生物试剂和生物制药的开发中发挥着重要作用。在这里，我们将主要回顾重组蛋白在包涵体中表达时的重折叠，尽管增强可溶性表达的策略被描述为重折叠包涵体的替代方法。这些策略包括，但不限于，在细胞培养基中添加化学伴侣，修改细胞裂解缓冲液和使用提高溶解度的融合标签。另一种增强溶解度的方法是为膜蛋白生成脂质复合物，形成不溶性的无脂蛋白。在各种增溶和再折叠技术中，介绍了使用变性剂、碱性pH值和压力的增溶和再折叠技术，重点介绍了使用洗涤剂的增溶和再折叠技术，这是一种有效且经济的技术。综述了十二烷基硫酸钠、月桂酰谷氨酸、萨科齐和十六烷基三甲基铵的广泛应用。缓慢或逐步去除用于增溶的变性剂或离子洗涤剂似乎在成功的再折叠中发挥关键作用，通过在再折叠过程中保持蛋白质的溶解度。在碱性再折叠中，缓慢的pH调节也有助于维持蛋白质的溶解度。加压折纸时，少量盐酸胍辅助折纸。

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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.