van Sommeren A P, Machielsen P A, Gribnau T C
AKZO, Pharma Division, Organon Teknika B.V. Clinical Lab. Systems Research Unit, The Netherlands.
Prep Biochem. 1992 Jun;22(2):135-49. doi: 10.1080/10826069208021364.
The binding capacity of protein A Sepharose 4 Fast Flow for mouse IgG1 monoclonal antibodies (mabs) appears to be highly dependent on the buffer composition with respect to both concentration and ion type. Depending on the particular mab dynamic binding capacities up to 20 mg mab per ml gel could be obtained, when these mabs were isolated from supernatants of protein free hollow fibre cell culture systems. Variation of linear flow rate from 10 up to 300 cm/h and temperature (4 degrees C versus 25 degrees C) had a slight effect on the dynamic binding capacity, when a high ionic strength buffer was used during adsorption. Applying optimum binding conditions, final IgG fractions with a purity of more than 95% monomeric IgG were obtained. However, as side effect of the use of binding buffers with high ionic strength, the binding of acid proteases was also promoted.
蛋白A琼脂糖4快速流动介质对小鼠IgG1单克隆抗体（mab）的结合能力似乎高度依赖于缓冲液的组成，包括浓度和离子类型。当从无蛋白中空纤维细胞培养系统的上清液中分离这些单克隆抗体时，根据特定的单克隆抗体，每毫升凝胶的动态结合能力可达20毫克单克隆抗体。在吸附过程中使用高离子强度缓冲液时，线性流速从10厘米/小时变化到300厘米/小时以及温度（4℃对25℃）对动态结合能力有轻微影响。应用最佳结合条件，可获得纯度超过95%的单体IgG的最终IgG级分。然而，作为使用高离子强度结合缓冲液的副作用，酸性蛋白酶的结合也会被促进。
