Abstract：
Purification of rAAV is a crucial unit operation of the AAV production process. It enables the capture of AAV and removal of contaminants such as host cell proteins, host cell DNA, and other cell culture-related impurities. Here we describe the purification of rAAV produced in insect cells Sf9/rBEV by immuno-affinity capture chromatography. The method is fully scale-amenable unlike other traditional purification methods based on ultracentrifugation. The method reported herein has two main steps: (1) the clarification of cell lysate by depth filtration and (2) the selective capture and single-step purification of AAV via immune-affinity chromatography. This purification method has been successfully implemented to purify the majority of wild-type AAV serotypes.
摘要：
rAAV的纯化是AAV生产过程的关键单元操作。它能够捕获AAV和去除污染物，如宿主细胞蛋白，宿主细胞DNA，和其他细胞培养相关的杂质。在这里，我们描述了通过免疫亲和捕获色谱法在昆虫细胞Sf9/rBEV中产生的rAAV的纯化。与基于超速离心的其他传统纯化方法不同，该方法完全适合规模。本文报道的方法具有两个主要步骤:(1)通过深度过滤澄清细胞裂解物和(2)通过免疫亲和层析选择性捕获和单步纯化AAV。该纯化方法已成功实施以纯化大多数野生型AAV血清型。
