Biogen, Gene Therapy Process Development, 225 Binney Street, Cambridge, MA, 02142, USA.
Biotechnol J. 2021 Jan;16(1):e2000015. doi: 10.1002/biot.202000015. Epub 2020 Oct 19.
The development of recombinant adeno-associated virus (rAAV) gene therapies is becoming an increasing priority in the biotherapeutic landscape. One of the challenges associated with the production of rAAV is the formation of empty AAV particles that do not contain a therapeutic gene. The concerns about the impact of empty particles on clinical safety and rAAV-mediated gene expression have necessitated the development of purification processes to remove these species. The development of a robust and scalable purification process to separate empty and full AAV particles at large scale remains a challenge. In this study, a novel anion exchange chromatography process based on isocratic wash and elution steps to enrich full rAAV2 particles is presented. An operating design space is identified to ensure the robustness of the process. The isocratic chromatography provides several advantages over the traditional shallow linear gradient elution, including lower buffer consumption, smaller intermediate pool volumes, and more robust manufacturing.
腺相关病毒（rAAV）基因治疗的发展正在成为生物治疗领域的一个日益优先事项。rAAV 生产面临的挑战之一是形成不包含治疗基因的空 AAV 颗粒。对空颗粒对临床安全性和 rAAV 介导的基因表达的影响的担忧，需要开发纯化工艺来去除这些物质。开发一种强大且可扩展的大规模分离空和满 AAV 颗粒的纯化工艺仍然是一个挑战。在这项研究中，提出了一种基于等度洗脱步骤的新型阴离子交换色谱工艺，用于富集完整的 rAAV2 颗粒。确定了一个操作设计空间，以确保工艺的稳健性。与传统的浅层线性梯度洗脱相比，等度色谱具有以下几个优点，包括更低的缓冲液消耗、更小的中间池体积和更稳健的制造。
