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.

中文翻译：

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使用等度阴离子交换色谱分离空的和完全重组的腺相关病毒颗粒

在生物治疗领域，重组腺相关病毒（rAAV）基因疗法的开发正变得越来越重要。与rAAV的产生相关的挑战之一是不包含治疗基因的空AAV颗粒的形成。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，使得必须开发去除这些物种的纯化方法。鲁棒且可扩展的纯化方法的发展，以大规模分离空的和完整的AAV颗粒仍然是一个挑战。在这项研究中，提出了一种基于等度洗涤和洗脱步骤以富集完整rAAV2颗粒的新型阴离子交换色谱方法。确定操作设计空间以确保过程的鲁棒性。