Winzor Donald J, Wills Peter R
Department of Biochemistry, School of Molecular and Microbial Sciences, University of Queensland, Brisbane, Qld 4072, Australia.
Biophys Chem. 2003 May 1;104(1):345-59. doi: 10.1016/s0301-4622(03)00003-6.
This investigation re-examines theoretical aspects of the allowance for effects of thermodynamic non-ideality on the characterization of protein self-association by frontal exclusion chromatography, and thereby provides methods of analysis with greater thermodynamic rigor than those used previously. Their application is illustrated by reappraisal of published exclusion chromatography data for hemoglobin on the controlled-pore-glass matrix CPG-120. The equilibrium constant of 100/M that is obtained for dimerization of the alpha(2)beta(2) species by this means is also deduced from re-examination of published studies of concentrated hemoglobin solutions by osmotic pressure and sedimentation equilibrium methods.
本研究重新审视了热力学非理想性对用前沿排阻色谱法表征蛋白质自缔合作用影响的理论方面，从而提供了比以前使用的方法具有更高热力学严谨性的分析方法。通过重新评估已发表的关于血红蛋白在可控孔径玻璃基质CPG - 120上的排阻色谱数据，说明了这些方法的应用。通过这种方法获得的α(2)β(2)物种二聚化的100/M的平衡常数，也是通过对已发表的用渗透压和沉降平衡方法研究浓缩血红蛋白溶液的研究重新审视推导出来的。
