Downstream Processing Group, Bioprocessing Technology Institute, Agency for Science, Technology and Research, Singapore, Singapore.
Cell Line Development Group, Bioprocessing Technology Institute, Agency for Science, Technology and Research, Singapore, Singapore.
PLoS One. 2023 Jan 25;18(1):e0280760. doi: 10.1371/journal.pone.0280760. eCollection 2023.
One of the key challenges in downstream bioprocessing is to obtain products of high purity in a productive fashion through the effective removal of process and product related impurities. While a classical simulated moving bed (SMB) system operation can typically achieve a 2-component separation between the weakly bound impurities and target species, here we present an advanced SMB approach that can achieve a 3-component separation, including the removal of the strongly bound impurities from the target species. As a proof-of-concept, we demonstrate the enhanced removal of strongly bound host cell proteins (HCP) from the target monoclonal antibody (mAb) through the utilisation of the advanced SMB approach in a non-affinity cation exchange (CEX) capture step. In this way, 1 less polishing step was required to achieve the therapeutic requirements of < 100 ppm HCP and the overall process recovery was increased by ~ 6% compared to the corresponding process that utilised a batch CEX operation. The non-affinity CEX capture platform technology established through the utilisation of the advanced SMB approach presented here can potentially be further applied to address the downstream processing challenges presented by other challenging biotherapeutic modalities to yield a final target product with improved purity and recovery.
下游生物加工的一个关键挑战是通过有效去除工艺和产品相关杂质，以高产率获得高纯度的产品。虽然经典的模拟移动床（SMB）系统操作通常可以实现弱结合杂质和目标物质之间的 2 组分分离，但在这里我们提出了一种先进的 SMB 方法，该方法可以实现 3 组分分离，包括从目标物质中去除强结合杂质。作为概念验证，我们通过在非亲和阳离子交换（CEX）捕获步骤中利用先进的 SMB 方法，从目标单克隆抗体（mAb）中增强去除强结合的宿主细胞蛋白（HCP）。这样，与使用批处理 CEX 操作的相应工艺相比，仅需 1 个额外的抛光步骤即可达到 <100ppm HCP 的治疗要求，并且整体工艺回收率提高了约 6%。通过利用此处提出的先进 SMB 方法建立的非亲和 CEX 捕获平台技术，有可能进一步应用于解决其他具有挑战性的生物治疗方式带来的下游加工挑战，以获得具有更高纯度和回收率的最终目标产品。
