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首页> 外文期刊>Biotechnology Journal: Healthcare,Nutrition,Technology >Systematic Interpolation Method Predicts Antibody Monomer-Dimer Separation by Gradient Elution Chromatography at High Protein Loads
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Systematic Interpolation Method Predicts Antibody Monomer-Dimer Separation by Gradient Elution Chromatography at High Protein Loads

机译：系统内插方法通过高蛋白质负荷通过梯度洗脱色谱法预测抗体单体二聚体分离

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摘要

A previously developed empirical interpolation (EI) method is extended to predict highly overloaded multicomponent elution behavior on a cation exchange (CEX) column based on batch isotherm data. Instead of a fully mechanistic model, the EI method employs an empirically modified multicomponent Langmuir equation to correlate two-component adsorption isotherm data at different salt concentrations. Piecewise cubic interpolating polynomials are then used to predict competitive binding at intermediate salt concentrations. The approach is tested for the separation of monoclonal antibody monomer and dimer mixtures by gradient elution on the cation exchange resin Nuvia HR-S. Adsorption isotherms are obtained over a range of salt concentrations with varying monomer and dimer concentrations. Coupled with a lumped kinetic model, the interpolated isotherms predict the column behavior for highly overloaded conditions. Predictions based on the EI method shows good agreement with experimental elution curves for protein loads up to 40 mg mL(-1) column or about 50% of the column binding capacity. The approach can be extended to other chromatographic modalities and to more than two components.
机译：先前开发的经验插值（EI）方法延伸以基于批量等温数据在阳离子交换（CEX）列上预测高度过载的多组分洗脱行为。 EI方法代替全机械模型，使用经验改性的多组分Langmuir方程，以在不同的盐浓度下关联双组分吸附等温数据。然后使用分段立方体内插多项式来预测中间盐浓度的竞争结合。通过阳离子交换树脂Nuvia HR-S对单克隆抗体单体和二聚体混合物进行单克隆抗体单体和二聚体混合物进行测试。在具有不同单体和二聚体浓度的一系列盐浓度上获得吸附等温物。耦合与集总动力学模型，内插等温物预测高度过载的条件的列行为。基于EI方法的预测显示了与蛋白质载荷高达40mg ml（-1）柱的实验洗脱曲线或柱结合能力的实验洗脱曲线的良好一致性。该方法可以扩展到其他色谱模式和两个以上的组件。

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