Thermodynamic modelling of hydrophobic interaction chromatography of biomolecules in the presence of salt

被引:11
|
作者
Mirani, Mohammad Reza [1 ]
Rahimpour, Farshad [1 ]
机构
[1] Razi Univ, Fac Engn, Dept Chem Engn, Biotechnol Res Lab, Kermanshah 6714967346, Iran
关键词
Hydrophobic interaction chromatography; Protein; Salt; Adsorption; Thermodynamic modelling; PROTEIN ADSORPTION; RETENTION; PURIFICATION; PRECIPITATION; EXTRACTION; MECHANISM; RECOVERY; SOLUTE;
D O I
10.1016/j.chroma.2015.10.019
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Hydrophobic interaction chromatography CHIC) is a useful method for isolation and purification of macromolecules. HIC separates proteins on the basis of surface hydrophobicity while generally retaining the activity of proteins. Aqueous mobile phases with high salt concentrations are often used to adsorb the proteins on a mildly hydrophobic support. In this research, the thermodynamic model of Chen and Sun, which predicts the adsorption isotherms of protein in presence of different type of salts, was modified by substitution the protein and salt activities in the mobile phase instead of their concentrations. In addition, model was examined for studying the adsorption of BSA, HSA, alpha-lactalbumin and Trypsinogen on different sepharose gels. The model parameters of Chen and Sun are adsorption equilibrium constant (K-p), protein dehydration equilibrium constant (K-s) salt coefficient (alpha) and number of ligand binding (n). By substitution activity instead of salt and protein concentration, two other parameters (c(1) and A(s)), which related to the activity coefficients, are added to the model. The parameters of this nonlinear model are calculated by genetic algorithm (GA). The maximum average absolute percentage deviation (AAD) for the data which are obtained from the adsorption isotherm of BSA on phenyl sepharose gel, in the presence of different concentration of NaCl was 4.8%, while for Chen and Sun model, was 22.0%. Also maximum ADD for FISA, alpha-lactalbumin, and Trypsinogen adsorption was 7.8, 6.9, and 8.4, respectively. The results indicate that the modified model has adequate accuracy to predict protein HIC behaviour. (C) 2015 Elsevier B.V. All rights reserved.
引用
收藏
页码:170 / 177
页数:8
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