Kang K A, Ryu D D
Department of Chemical Engineering, University of California, Davis 95616.
Biotechnol Prog. 1991 May-Jun;7(3):205-12. doi: 10.1021/bp00009a002.
Effects of operational and system parameters on process scale-up of murine immunoglobulin (IgG2a kappa) purification using Protein A affinity chromatography are investigated. Parameters studied are those related to sample application, elution, ligand concentration on support, and column size change. Between sample application velocities of 0.004 and 0.030 cm/s (16-108 cm/h), the product concentration profiles in eluate did not show significant differences. With given system parameters, the retention time and bandwidth of the peak could be predicted by moment theory. The mean equilibrium constant during elution showed a strong effect of pH between 2 and 4. Within the range of protein A concentrations studied, 0.15-1.22 mg/mL of gel, the column capacity shows a linear relationship with the concentration of protein A immobilized. Dimensional scale-up of the column in the radial direction increased the total purification capacity linearly as the cross-sectional area increased without increasing pressure drop; however, the product concentration was diluted. Scale-up of the column dimension in the axial direction enables higher concentrations of product in the eluate, although the retention time increases linearly as the gel height increases.
研究了操作参数和系统参数对使用蛋白A亲和色谱法扩大小鼠免疫球蛋白（IgG2a κ）纯化工艺规模的影响。所研究的参数包括与上样、洗脱、配体在载体上的浓度以及柱尺寸变化相关的参数。在上样速度为0.004至0.030 cm/s（16 - 108 cm/h）之间时，洗脱液中产物浓度曲线未显示出显著差异。在给定的系统参数下，峰的保留时间和带宽可以通过矩理论进行预测。洗脱过程中的平均平衡常数在pH值为2至4之间时显示出强烈的影响。在所研究的蛋白A浓度范围内，即凝胶中蛋白A浓度为0.15 - 1.22 mg/mL，柱容量与固定化蛋白A的浓度呈线性关系。柱在径向方向上的尺寸扩大随着横截面积的增加线性增加了总纯化能力，且不增加压降；然而，产物浓度被稀释了。柱在轴向方向上的尺寸扩大能够使洗脱液中产物浓度更高，尽管保留时间随着凝胶高度的增加而线性增加。
