Harlan J E, Picot D, Loll P J, Garavito R M
Department of Biochemistry and Molecular Biology, University of Chicago, Illinois 60637, USA.
Anal Biochem. 1995 Jan 20;224(2):557-63. doi: 10.1006/abio.1995.1087.
A method for the calibration of size-exclusion chromatographic columns is proposed that takes into account the nonlinear dependence of the Stokes radius Rs upon the partition coefficient KD. The method is based on the assumption that the pore size distribution of a molecular sieve column can be described by the sum of two Gaussian distributions and has been successfully tested with low-pressure chromatography gels (Sephacryl and Superose) and HPLC gels (TSK SW) over a wide range of protein sizes. An application of this method is described, in which aggregation states of the membrane protein prostaglandin H2 synthase solubilized in nonionic detergents are estimated.
提出了一种尺寸排阻色谱柱的校准方法，该方法考虑了斯托克斯半径Rs对分配系数KD的非线性依赖性。该方法基于这样的假设，即分子筛柱的孔径分布可以用两个高斯分布的总和来描述，并且已经在广泛的蛋白质大小范围内，用低压色谱凝胶（Sephacryl和Superose）和HPLC凝胶（TSK SW）成功进行了测试。描述了该方法的一个应用，其中估计了溶解在非离子洗涤剂中的膜蛋白前列腺素H2合酶的聚集状态。
