Department of Chemical Engineering, McMaster University, 1280 Main Street West, Hamilton, Ontario L8S 4L7, Canada.
Department of Chemical Engineering, McMaster University, 1280 Main Street West, Hamilton, Ontario L8S 4L7, Canada.
J Chromatogr B Analyt Technol Biomed Life Sci. 2018 Jun 1;1086:23-28. doi: 10.1016/j.jchromb.2018.04.002. Epub 2018 Apr 4.
In recent papers we have proposed cuboid packed-bed devices as viable alternatives to preparative columns with low bed-height to diameter ratios that are typically used for biopharmaceutical purification processes. In this case study, we systematically examined operating parameters such as mobile phase flow rate, sample injection volume and feed protein concentration on the characteristics of flow-through and bound-and-eluted protein peaks. The current study was carried out using strong anion exchange media packed in a cuboid packed-bed device and its equivalent column, i.e. with identical volume and bed-height. Our experimental results showed that the cuboid packed-bed device outperformed its equivalent column at all conditions examined. However, the improvement in performance in flow-through experiments was more significant at lower flow rates, and when smaller sample volumes (i.e. less than 20% of bed volume) were injected. Also, the performance of the cuboid packed-bed was significantly better when a concentrated protein sample was injected using a small volume loop. In bind-and-elute experiments, the flow rate had a very significant impact on the performance of the cuboid packed-bed device, with better results being obtained at the lower flow rates examined. By choosing appropriate experimental conditions, significantly sharper peaks and thereby efficient separations could be achieved using the cuboid packed-bed device than with its equivalent column.
在最近的论文中，我们提出了使用长方体填充床设备作为替代方案，替代通常用于生物制药纯化过程的低床高径比的制备柱。在本案例研究中，我们系统地研究了流动相流速、样品进样量和进料蛋白浓度等操作参数对流动相和结合洗脱蛋白峰特征的影响。本研究使用强阴离子交换介质填充在长方体填充床设备及其等效柱中进行，即具有相同的体积和床高。我们的实验结果表明，在所有考察的条件下，长方体填充床设备的性能均优于其等效柱。然而，在较低流速和较小样品体积（即小于床体积的 20%）进样时，流动相实验中的性能改善更为显著。此外，当使用小体积环注入浓缩蛋白样品时，长方体填充床的性能明显更好。在结合洗脱实验中，流速对长方体填充床设备的性能有很大的影响，在考察的较低流速下获得了更好的结果。通过选择适当的实验条件，使用长方体填充床设备可以比使用其等效柱获得更尖锐的峰，从而实现更有效的分离。
