化学
盐析
亲水作用色谱法
聚乙二醇
色谱法
PEG比率
盐(化学)
疏水效应
两亲性
脱水
离子色谱法
有机化学
生物化学
高效液相色谱法
水溶液
共聚物
经济
聚合物
财务
作者
Tsutomu Arakawa,Yui Tomioka,Yasunori Kurosawa,Teruo Akuta
标识
DOI:10.1016/j.chroma.2023.464091
摘要
Hydrophobic interaction chromatography (HIC) is a commonly used chromatography technique for purifying proteins. It utilizes salting-out salts to facilitate the binding of native proteins to weakly hydrophobic ligands. There have been three proposed mechanisms for the promoting effects of salting-out salts, which include the dehydration of proteins by salts, cavity theory, and salt exclusion. To evaluate the above three mechanisms, an HIC study was conducted on Phenyl Sepharose using four different additives. These additives included a salting-out salt (NH4)2SO4, sodium phosphate that increases the surface tension of water, a salting-in salt MgCl2, and an amphiphilic protein-precipitant polyethylene glycol (PEG). Results indicated that the first two salts resulted in protein binding, while MgCl2 and PEG led to flow-through. These findings were then used to interpret the three proposed mechanisms, which showed that MgCl2 and PEG deviated from the dehydration mechanism, and MgCl2 also deviated from the cavity theory. The observed effects of these additives on HIC were reasonably well explained for the first time by their interactions with proteins.
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