Graduate School of Science and Technology for Innovation, Yamaguchi University, Ube, Japan.
Biomedical Engineering Center (YUBEC), Yamaguchi University, Ube, Japan.
Biotechnol Bioeng. 2021 Mar;118(3):1262-1272. doi: 10.1002/bit.27649. Epub 2021 Jan 3.
A new method was proposed for increasing the capture chromatography process efficiency, linear flow-velocity gradient (LFG). The method uses a linear decreasing flow-velocity gradient with time during the sample loading. The initial flow velocity, the final flow velocity and the gradient time are the parameters to be tuned. We have developed a method for determining these parameters by using the total column capacity and the total loaded amount as a function of time. The capacity can be calculated by using the relationships between dynamic binding capacity (DBC) and residence time. By leveraging the capacity, loading amount, and the required conditions, the optimum LFG can be designed. The method was verified by ion-exchange and protein A chromatography of monoclonal antibodies (mAbs). A two-fold increase in the productivity during the sample loading was possible by LFG compared with the constant flow-velocity (CF) operation. LFG was also applied to a 4-column continuous process. The simulation showed that the cost of resin per unit amount of processed mAbs can be reduced by 13% while 1.4 times enhancement in productivity was preserved after optimization by LFG compared to CF. The process efficiency improvement is more pronounced when the isotherm is highly favorable and the loading volume is large.
提出了一种提高捕获色谱过程效率的新方法，即线性流速梯度（LFG）。该方法在样品加载过程中随时间采用线性降低的流速梯度。初始流速、最终流速和梯度时间是需要调整的参数。我们已经开发了一种方法，通过使用总柱容量和总加载量作为时间的函数来确定这些参数。容量可以通过动态结合容量（DBC）和停留时间之间的关系来计算。通过利用容量、加载量和所需条件，可以设计出最佳的 LFG。该方法通过单克隆抗体（mAbs）的离子交换和蛋白 A 色谱进行了验证。与恒流速（CF）操作相比，LFG 可在样品加载过程中将生产率提高两倍。LFG 还应用于 4 柱连续工艺。模拟表明，通过 LFG 优化后，与 CF 相比，每单位处理 mAbs 的树脂成本可降低 13%，同时保持 1.4 倍的生产率提高。当等温线非常有利且加载体积较大时，过程效率的提高更为显著。
