Bioprocess Engineering Group, Max Planck Institute for Dynamics of Complex Technical Systems, Sandtorstr. 1, 39106 Magdeburg, Germany; Sartorius Stedim Biotech GmbH, August-Spindler Strasse 11, 37079 Göttingen, Germany.
Sartorius Stedim Biotech GmbH, August-Spindler Strasse 11, 37079 Göttingen, Germany.
J Biotechnol. 2021 Dec 10;342:139-148. doi: 10.1016/j.jbiotec.2021.10.003. Epub 2021 Oct 20.
Robust and flexible continuous unit operations that enable the establishment of intensified bioprocesses is one of the most relevant trends in manufacturing of biopharmaceuticals, including virus-based products. Sulfated cellulose membrane adsorbers (SCMA) are one of the most promising matrices for chromatographic purification of virus particles, like influenza viruses. Here, a three 'column' periodical counter current set-up was used to continuously purify influenza A/PR/8/34 virus particles using SCMA in bind-elute mode. It was possible to recover 67.4% of the HA-activity and to remove 67.4% and 99.8% of the total protein and DNA, respectively. The performance of the continuous process operated over a total of 10 loops, was slightly inferior to was obtained in a comparable batch process. Nevertheless, it was possible to increase the effective usage of binding capacity to 80%, resulting on a productivity of 22.8 kHAU ml min. As a proof-of-principle, SCMA were successfully used as matrix for purification of cell-derived influenza virus particles, in continuous mode.
建立强化生物工艺的稳健且灵活的连续单元操作是生物制药（包括基于病毒的产品）制造中最相关的趋势之一。硫酸化纤维素膜吸附剂（SCMA）是用于病毒颗粒（如流感病毒）色谱纯化的最有前途的基质之一。在这里，使用三列周期性逆流装置以绑定洗脱模式连续纯化流感 A/PR/8/34 病毒颗粒。可以回收 67.4%的 HA 活性，分别去除 67.4%和 99.8%的总蛋白和 DNA。在总共 10 个循环中运行的连续工艺的性能略低于可比批次工艺的性能。然而，有可能将结合容量的有效利用率提高到 80%，从而使生产率达到 22.8 kHAU ml min。作为原理验证，SCMA 成功地用作连续模式下细胞来源的流感病毒颗粒的纯化基质。
