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A model for the salt effect on adsorption equilibrium of basic protein to dye-ligand affinity adsorbent

机译：盐对碱性蛋白对染料-配体亲和吸附剂吸附平衡的影响模型

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摘要

A model describing the salt effect on adsorption equilibrium of a basic protein, lysozyme, to Cibacron Blue 3GA-modified Sepharose CL-6B (CB-Sepharose) has been developed. In this model, it is assumed that the presence of salt causes a fraction of dye-ligand molecules to lodge to the surface of the agarose gel, resulting from the induced strong hydrophobic interaction between dye ligand and agarose matrix. The salt effect on the lodging of dye-ligand is expressed by the equilibrium between salt and dye-ligand. For the interactions between protein and vacant binding sites, stoichiometric equations based either on cation exchanges or on hydrophobic interactions are proposed since the CB dye can be regarded as a cation exchanger contributed by the sulfonate groups on it. Combining with the basic concept of steric mass-action theory for ion exchange, which considers both the multipoint nature and the macromolecular steric shielding of protein adsorption, an explicit isotherm for protein adsorption equilibrium on the dye-ligand adsorbent is formulated, involving salt concentration as a variable. Analysis of the model parameters has yielded better understanding of the mechanism of salt effects on adsorption of the basic protein. Moreover, the model predictions are in good agreement with the experimental data over a wide range of salt and ligand concentrations, indicating the predictive nature of the model.
机译：建立了描述盐对碱性蛋白溶菌酶对Cibacron Blue 3GA修饰的Sepharose CL-6B（CB-Sepharose）吸附平衡的影响的模型。在该模型中，假定盐的存在会导致一部分染料-配体分子滞留在琼脂糖凝胶表面，这是由于染料配体和琼脂糖基质之间产生了强烈的疏水性相互作用所致。盐对染料-配体的沉降的作用通过盐和染料-配体之间的平衡来表达。对于蛋白质和空的结合位点之间的相互作用，提出了基于阳离子交换或基于疏水相互作用的化学计量方程式，因为CB染料可被认为是由其上的磺酸根基团贡献的阳离子交换剂。结合空间质量作用理论中离子交换的基本概念，它考虑了蛋白质吸附的多点性质和大分子空间屏蔽作用，制定了一个明确的等温线用于染料-配体吸附剂上的蛋白质吸附平衡，其中盐浓度为一个变量。模型参数的分析已使人们更好地了解了盐对碱性蛋白吸附的影响机理。此外，模型预测值与各种盐和配体浓度范围内的实验数据高度吻合，表明了模型的预测性质。

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