折叠前的净化程序,以包含来自身体的非标记的阴离子重组蛋白质,具有多重二硫化物键,以进行高效的重新折叠
Shuichiro Kimura1, Wataru Yamamoto2, Ai Miyamoto2
1Division of Applied Chemistry, Graduate School of Natural Science and Technology, Okayama University, Okayama, Japan.
Biotechnology progress
|January 26, 2025
概括
这项研究引入了一种使用可逆S-化处理的新型重叠前净化方法. 这种技术通过从包含体中去除杂质和降解产物来提高重组蛋白重新折叠产量.
科学领域:
- 生物化学 生物化学
- 蛋白质化学 蛋白质化学
- 生物技术是生物技术.
背景情况:
- 含有二硫化物的重组蛋白通常需要从包含体重新折叠.
- 包括体中的杂质阻碍了有效的重新折叠和净化.
- 要去除抑制剂,需要进行重新折叠前的关键净化步骤.
研究的目的:
- 开发一种新的重新折叠前净化程序,以改善重组蛋白质生产.
- 通过有效地去除污染物和降解产品来提高重新折叠产量.
- 提出一种无标签净化方法,简化下游加工.
主要方法:
- 使用可逆S-化技术进行蛋白质溶解.
- 使用逆相高性能液态染色学 (RP-HPLC) 进行净化.
- 将该方法应用于一种高度阴性小鼠血管内皮细胞生长因子 (mVEGF) 模型蛋白.
主要成果:
- 能够有效地去除再折叠抑制剂和蛋白质分解性降解产品.
- 与传统方法相比,证明了改进的重新折叠产量.
- 成功净化了一种含有多硫化物的同位体mVEGF蛋白 (pI = 9.25).
结论:
- 可逆S-化技术提供了一种优越的重新折叠前净化策略.
- 这种无标签的方法简化了下游处理,消除了需要移除亲和标签的需求.
- 开发的程序是有效的生产复杂的二硫化物含有复合蛋白质,如mVEGF.
相关概念视频
Protein Folding
7.7K
Proteins are chains of amino acids linked together by peptide bonds. Upon synthesis, a protein folds into a three-dimensional conformation, critical to its biological function. Interactions between its constituent amino acids guide protein folding, and hence the protein structure is primarily dependent on its amino acid sequence.
Protein Structure Is Critical to Its Biological Function
Proteins perform a wide range of biological functions such as catalyzing chemical reactions, providing...
Protein Structure Is Critical to Its Biological Function
Proteins perform a wide range of biological functions such as catalyzing chemical reactions, providing...
7.7K
Molecular Chaperones and Protein Folding
17.7K
The native conformation of a protein is formed by interactions between the side chains of its constituent amino acids. When the amino acids cannot form these interactions, the protein cannot fold by itself and needs chaperones. Notably, chaperones do not relay any additional information required for the folding of polypeptides; the native conformation of a protein is determined solely by its amino acid sequence. Chaperones catalyze protein folding without being a part of the folded protein.
The...
The...
17.7K


