Protein partition between the different phases comprising poly(ethylene glycol)-salt aqueous two-phase systems, hydrophobic interaction chromatography and precipitation: A generic description in terms of salting-out effects

被引:41
作者
Huddleston, J
Abelaira, JC
Wang, RD
Lyddiatt, A
机构
[1] Biochemical Recovery Group, School of Chemical Engineering, University of Birmingham
来源
JOURNAL OF CHROMATOGRAPHY B-BIOMEDICAL APPLICATIONS | 1996年 / 680卷 / 1-2期
关键词
partitioning; aqueous two-phase systems; salting-out; proteins; polyethylene glycol;
D O I
10.1016/0378-4347(95)00448-3
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The solution behaviour of selected proteins has been studied under conditions promoting precipitation, binding to mildly hydrophobic adsorbents or partition. Solvophobic theory may be used to describe these forms of protein partition. The tendency of a protein to partition therein is dependent upon surface properties of the protein solute mediated by the concentration and nature of added salts. As applied to partitioning in poly(ethylene glycol) (PEG)-salt systems this implies that linear (Bronsted) relationships apply only to proteins partitioned close to the critical point. At longer tie-line lengths protein partitioning is increasingly influenced by salting-out forces. This is confirmed by the observed behaviour of the proteins. The point at which this behaviour changes has been unambiguously defined enabling the direct comparison of phase transition of proteins during partition in all systems. The results obtained show that phase transition during adsorption and partition occur at similar concentrations of salt. This is less than that required to promote precipitation. It appears, from these limited studies, that top-phase preferring proteins are partitioned at salt concentrations above those required to cause adsorption. Proteins preferring the lower phase are partitioned at salt concentrations close to or below those required for adsorption. This raises questions regarding the solvated molecular form of the partitioned proteins and the definition of the partition coefficient.
引用
收藏
页码:31 / 41
页数:11
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