可扩展的双列阴离子交换亲和力色谱基于平台的过程,用于重组蛋白质净化
Sai Vivek Prabhala1, David W Wood1
1William G. Lowrie Department of Chemical and Biomolecular Engineering, The Ohio State University, 460C CBEC Building, 151 W. Woodruff Ave., Columbus, OH, 43210, USA.
Protein expression and purification
|February 9, 2024
概括
一种新的双列净化方法使用氨酸结合标签进行初始捕获,并使用iCapTag去除标签,产生纯粹的,无标签的重组蛋白质. 这种可扩展的平台提高了蛋白质净化效率和纯度.
科学领域:
- 生物化学 生物化学
- 蛋白质净化 蛋白质净化
- 生物技术是生物技术.
背景情况:
- 重组蛋白净化对于研究和治疗至关重要.
- 现有的方法经常面临纯度,标签删除或可扩展性的挑战.
- 需要新的亲和标签和净化策略来克服这些局限性.
研究的目的:
- 开发和验证用于重组蛋白质的新型双列净化策略.
- 为了证明结合肝素结合标签 (HB-tag) 和自删除iCapTagTM系统的有效性.
- 为获得高纯度,无标签蛋白质提供一个可扩展的平台.
主要方法:
- 设计了一种双列染色学工艺,使用阴离子交换和iCapTagTM 分裂-intein亲和树脂.
- 蛋白质最初被捕获使用HB标签在一个离子交换树脂.
- 随后的净化和标签去除使用iCapTagTM系统实现,并进行一夜间化.
主要成果:
- 双列方法成功净化了各种蛋白质,包括eGFP,βgal,MBP和βlac.
- 实现了高纯度水平和有效的宿主细胞DNA清除.
- 该过程证明了可扩展性和高效的标签自删除.
结论:
- 拟议的双联亲和标签系统为重组蛋白净化提供了一个多功能和高效的平台.
- 这种方法结合了离子交换能力和亲和标签选择性的好处.
- 该技术可以生产适合各种下游应用的无标签蛋白质.
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