使用SO3单质色谱对rAAV捕获阶段净化进行优化
Kaja Bažec1, Mirjam Krašna1, Andrej Mihevc1
1Process Development for Viral Vectors and Vaccines, Sartorius BIA Separations, Ajdovščina, Slovenia.
Electrophoresis
|August 21, 2023
概括
这项研究优化了使用阴离子交换色谱的腺相关病毒 (AAV) 净化. 优化的过程实现了高载体恢复和显著减少蛋白质和DNA杂质,用于基因疗法应用.
科学领域:
- 生物技术是生物技术.
- 基因治疗 基因治疗
- 下游加工 下游加工
背景情况:
- 腺相关病毒 (AAV) 载体对于基因疗法至关重要.
- 严格的纯度是需要在体内给予AV的.
- 下游加工必须遵守监管指南.
研究的目的:
- 为了优化非亲和性,将重组AAV (rAAV) 捕获的血清型独立.
- 为了评估对流相互作用介质 (CIM) 离子交换SO3单体,用于RAAV捕获.
- 为了确定最佳的绑定移动相条件,以捕获rAAV.
主要方法:
- 使用CIM SO3 96井板选缓冲 pH,NaCl 度和波洛克萨默 188 的选.
- 从Sf9细胞系中分别预处理的病毒样本的分析.
- 确定动态结合能力,并在CIMmultus工业线上进行重新测试.
主要成果:
- 实现了51%的高整体载体恢复.
- 显著减少杂质:蛋白质的99.98%和DNA的99.25%.
- 优化的捕获步骤参数在工业规模上得到证实.
结论:
- 阳离子交换SO3单体捕获步骤已成功优化.
- 这种优化增强了用于基因治疗的rAAV的下游处理.
- 开发的方法符合在体内应用的严格纯度要求.
相关概念视频
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