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阐明树脂介质,料成分和工艺条件之间的相互作用在AAV亲和系列中,用同焦显微镜进行染

Harshal Soni^1,2, Sergey Pryshchep², Thomas F Johnson³

¹Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Troy, New York, USA.

Biotechnology and bioengineering

|September 10, 2025

概括

同焦点显微镜显示了腺相关病毒 (AAV) 树脂载荷和污染的差异. 基于PSDVB的树脂最初显示出完全的访问,但在重复使用时会受到污染,与基于阿加的树脂不同.

科学领域:

生物技术是生物技术.
染色体学染色体学是一种染色学.
显微镜的使用方法

背景情况:

腺相关病毒 (AAV) 净化依赖于亲和性染色学.
在加载,化和清洗过程中了解树脂的行为对于过程优化至关重要.
树脂再利用影响生物制药制造中的效率和纯度.

研究的目的:

通过共聚焦显微镜来研究腺相关病毒 (AAV) 在亲和树脂中的加载和污染.
为了比较基于PSDVB的POROS CaptureSelect (PCS) 和基于agarose的AVIPure AAV9树脂的性能和可重复使用性.
阐明树脂特性和工艺条件对柱子寿命的影响.

主要方法:

使用共聚焦显微镜,在各种工艺步骤后检查树脂珠中的AAV吸附概况.
商用AAV亲和树脂 (PCS和AVIPure) 用不同的料混合物进行了测试.
使用高分辨率的X射线CT扫描来分析树脂毛孔结构.

主要成果:

最初,PCS树脂允许AAV完全访问,而AVIPure树脂显示表面限制的结合.
由于AAV和杂质的携带,在重复使用时发生了PCS树脂污染,导致孔隙封闭.
在清洁现场 (CIP) 过程中,AVIPure树脂证明了有效的AAV化和去除.

结论:

树脂毛孔大小 (PCS大毛孔) 影响了最初的AAV运输.
重复使用后PCS树脂中的污染严重阻碍了后续循环中的溶液运输.
在测试条件下,与PCS树脂相比,AVIPure树脂具有更好的可重复使用性和抗污染性.

关键词:

一个X射线CTCT.腺相关病毒 (AAV)亲缘树脂是一种亲缘树脂.染色体学染色体学染色体学柱子中的污染.共聚焦激光扫描显微镜 (CLSM)影像成像技术影像成像技术

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