da Silva Nuno R, Ferreira Luisa A, Madeira Pedro P, Teixeira José A, Uversky Vladimir N, Zaslavsky Boris Y
IBB-Institute for Biotechnology and Bioengineering, Centre of Biological Engineering, Universidade do Minho, Campus de Gualtar, 4710-057 Braga, Portugal; Institute for Biological Instrumentation, Russian Academy of Sciences, Pushchino, Moscow Region 142290, Russia.
Analiza, Inc., 3615 Superior Ave., Cleveland, OH 44114, USA; Institute for Biological Instrumentation, Russian Academy of Sciences, Pushchino, Moscow Region 142290, Russia.
J Chromatogr A. 2015 Oct 9;1415:1-10. doi: 10.1016/j.chroma.2015.08.053. Epub 2015 Aug 28.
Partition behavior of nine small organic compounds and six proteins was examined in poly(ethylene glycol)-8000-sodium sulfate aqueous two-phase systems containing 0.5M osmolyte (sorbitol, sucrose, trehalose, TMAO) and poly(ethylene glycol)-10000-sodium sulfate system, all in 0.01M sodium phosphate buffer, pH 6.8. The differences between the solvent properties of the coexisting phases (solvent dipolarity/polarizability, hydrogen bond donor acidity, and hydrogen bond acceptor basicity) were characterized with solvatochromic dyes using the solvatochromic comparison method. Differences between the electrostatic properties of the phases were determined by analysis of partitioning of sodium salts of dinitrophenylated (DNP-) amino acids with aliphatic alkyl side-chain. It was found out that the partition coefficient of all compounds examined (including proteins) may be described in terms of solute-solvent interactions. The results obtained in the study show that solute-solvent interactions of nonionic organic compounds and proteins in polyethylene glycol-sodium sulfate aqueous two-phase system differ from those in polyethylene glycol-dextran system.
在含有0.5M渗透剂(山梨醇、蔗糖、海藻糖、三甲胺氧化物)的聚乙二醇-8000-硫酸钠水两相体系以及聚乙二醇-10000-硫酸钠体系中,于0.01M磷酸钠缓冲液(pH 6.8)中研究了9种小有机化合物和6种蛋白质的分配行为。使用溶剂化显色比较法,通过溶剂化显色染料对共存相的溶剂性质差异(溶剂偶极矩/极化率、氢键给体酸度和氢键受体碱度)进行了表征。通过分析具有脂肪族烷基侧链的二硝基苯基化(DNP-)氨基酸钠盐的分配情况,确定了各相的静电性质差异。结果发现,所有检测化合物(包括蛋白质)的分配系数都可以用溶质-溶剂相互作用来描述。该研究所得结果表明,聚乙二醇-硫酸钠水两相体系中,非离子有机化合物和蛋白质的溶质-溶剂相互作用与聚乙二醇-葡聚糖体系中的不同。