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凝胶过滤色谱法测定大分子斯托克斯半径的解读

Interpretation of the stokes radius of macromolecules determined by gel filtration chromatography.

作者信息

Horiike K, Tojo H, Yamano T, Nozaki M

出版信息

J Biochem. 1983 Jan;93(1):99-106. doi: 10.1093/oxfordjournals.jbchem.a134183.

Abstract

From a comparison of the gel chromatographic properties of large randomly-coiled polypeptides in 6 M guanidine hydrochloride and of large globular proteins, we found that the distribution coefficient was more closely correlated with the intrinsic viscosity-based Stokes radius than with the translational frictional coefficient-based Stokes radius. This means that the effect of the hydrodynamic flow of dissolved molecules during gel chromatography should be considered. The ratio of transport of solute by bulk flow as compared with that by net diffusion (i.e., Brownian motion) is large under some conditions. On the other hand, we consider that the distribution coefficient obtained in static equilibrium experiments should be determined by the translational frictional coefficient-based Stokes radius, since the solvent does not flow. On this basis, we discuss the meaning of the Stokes radius and the separation mechanism of macromolecules by gel filtration.

摘要

通过比较大的无规卷曲多肽在6M盐酸胍中的凝胶色谱性质与大的球状蛋白质的凝胶色谱性质,我们发现分配系数与基于特性粘度的斯托克斯半径的相关性比与基于平移摩擦系数的斯托克斯半径的相关性更紧密。这意味着在凝胶色谱过程中应考虑溶解分子的流体动力学流动的影响。在某些条件下,溶质通过总体流动的传输与通过净扩散(即布朗运动)的传输之比很大。另一方面,我们认为在静态平衡实验中获得的分配系数应由基于平移摩擦系数的斯托克斯半径决定,因为溶剂不流动。在此基础上,我们讨论了斯托克斯半径的意义以及凝胶过滤分离大分子的机制。

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