Xiao Yunzhi, Rathore Anurag, O'Connell John P, Fernandez Erik J
Department of Chemical Engineering, University of Virginia, 102 Engineers' Way, Charlottesville, VA 22904-4741, USA.
J Chromatogr A. 2007 Jul 20;1157(1-2):197-206. doi: 10.1016/j.chroma.2007.05.009. Epub 2007 May 6.
A two-conformation adsorption model that includes the effects of salt concentration and temperature on both stability and adsorption has been developed to describe the effects of secondary protein unfolding on hydrophobic interaction chromatography (HIC). The model has been applied to a biotech protein and to beta-lactoglobulin on Phenyl Sepharose 6FF low sub HIC media. Thermodynamic property models for adsorption and protein stability with parameters obtained from experimental chromatographic data successfully describe observed chromatographic behavior over ranges of temperature and salt concentration, provide predictions of distribution among different conformers, and give a basis for calculating trends in retention strength and stability with changing conditions, that might prove useful in HIC process development.
已开发出一种双构象吸附模型，该模型考虑了盐浓度和温度对稳定性及吸附的影响，用于描述二级蛋白质解折叠对疏水作用色谱（HIC）的影响。该模型已应用于一种生物技术蛋白以及苯基琼脂糖6FF低取代度HIC介质上的β-乳球蛋白。利用从实验色谱数据获得的参数建立的吸附和蛋白质稳定性热力学性质模型，成功地描述了在温度和盐浓度范围内观察到的色谱行为，预测了不同构象之间的分布，并为计算随条件变化的保留强度和稳定性趋势提供了依据，这可能对HIC工艺开发有用。
