Tubio Gisela, Nerli Bibiana, Picó Guillermo
Physico-Chemistry Department, Faculty of Biochemical and Pharmaceutical Sciences and CONICET, National University of Rosario, Rosario 2000, Argentina.
J Chromatogr B Analyt Technol Biomed Life Sci. 2004 Jan 25;799(2):293-301. doi: 10.1016/j.jchromb.2003.10.060.
In order to develop possible correlations to predict partioning behaviour of proteins, five mammalian albumins (goat, bovine, equine, human and pig ones) with similar physico-chemical properties (molecular mass and isoelectrical point) were chosen. Evaluation of the relationship between hydrophobicity and partitioning coefficient (Kr) in polyethylenglycol-dextran (PEG-DxT500) systems formed by polyethyleneglycols of different molecular mass (3350, 6000 and 10,000) was investigated by estimating relative surface hydrophobicity (So) with a fluorescent probe, 1 anilino-8-naphthalene sulfonate. No relationship between Kr and So was found for systems formed by PEG3350, while aqueous two-phase systems with PEG6000 and PEG10,000 gave better correlations. The results obtained may be explained on the basis of an increase in the interaction between the latter PEGs and the protein due to their higher hydrophobic character which increases as the PEG molecular mass does so. In this way, systems with PEGs of higher molecular mass give the highest resolution to exploit hydrophobicity in partitioning.
为了建立可能的相关性以预测蛋白质的分配行为，选择了五种具有相似物理化学性质（分子量和等电点）的哺乳动物白蛋白（山羊、牛、马、人及猪的白蛋白）。通过用荧光探针1-苯胺基-8-萘磺酸盐估算相对表面疏水性（So），研究了在由不同分子量（3350、6000和10000）的聚乙二醇形成的聚乙二醇-葡聚糖（PEG-DxT500）体系中疏水性与分配系数（Kr）之间的关系。对于由PEG3350形成的体系，未发现Kr与So之间存在关系，而含有PEG6000和PEG10000的双水相体系具有更好的相关性。所得结果可以基于后两种聚乙二醇与蛋白质之间相互作用的增加来解释，这是由于它们较高的疏水特性随着聚乙二醇分子量的增加而增强。这样，具有较高分子量聚乙二醇的体系在分配中利用疏水性时具有最高的分辨率。
