公开(公告)号：US09198984B2

公开(公告)日：2015-12-01

申请号：US12226588

申请日：2007-04-27

Applicant: Martin Lock ,  Luc H Vandenberghe ,  James M. Wilson

Inventor： Martin Lock ,  Luc H Vandenberghe ,  James M. Wilson

IPC: A61K38/00 ,  C12N7/00 ,  A61K48/00 ,  C07K14/005

CPC classification number: C12N7/00 ,  A61K48/0091 ,  C07K14/005 ,  C12N2750/14151

Abstract: A method for producing AAV, without requiring cell lysis, is described. The method involves harvesting AAV from the supernatant. For AAV having capsids with a heparin binding site, the method involves modifying the AAV capsids and/or the culture conditions to ablate the binding between the AAV heparin binding site and the cells, thereby allowing the AAV to pass into the supernatant, i.e., media. Thus, the method of the invention provides supernatant containing high yields of AAV which have a higher degree of purity from cell membranes and intracellular materials, as compared to AAV produced using methods using a cell lysis step.

Abstract translation: 描述了不需要细胞裂解的生产AAV的方法。 该方法包括从上清液中收获AAV。 对于具有肝素结合位点的衣壳的AAV，该方法包括修饰AAV衣壳和/或培养条件以消除AAV肝素结合位点与细胞之间的结合，从而允许AAV进入上清液，即培养基 。 因此，与使用细胞裂解步骤的方法产生的AAV相比，本发明的方法提供了含有高产率的AAV的上清液，其具有比细胞膜和细胞内物质更高的纯度。

公开(公告)号：US20090275107A1

公开(公告)日：2009-11-05

申请号：US12226588

申请日：2007-04-27

Applicant: Martin Lock ,  Luc H. Vandenberghe ,  James M. Wilson

Inventor： Martin Lock ,  Luc H. Vandenberghe ,  James M. Wilson

IPC: C12N7/02 ,  C12N7/00

CPC classification number: C12N7/00 ,  A61K48/0091 ,  C07K14/005 ,  C12N2750/14151

Abstract: A method for producing AAV, without requiring cell lysis, is described. The method involves harvesting AAV from the supernatant. For AAV having capsids with a heparin binding site, the method involves modifying the AAV capsids and/or the culture conditions to ablate the binding between the AAV heparin binding site and the cells, thereby allowing the AAV to pass into the supernatant, i.e., media. Thus, the method of the invention provides supernatant containing high yields of AAV which have a higher degree of purity from cell membranes and intracellular materials, as compared to AAV produced using methods using a cell lysis step.

Abstract translation: 描述了不需要细胞裂解的生产AAV的方法。 该方法包括从上清液中收获AAV。 对于具有肝素结合位点的衣壳的AAV，该方法包括修饰AAV衣壳和/或培养条件以消除AAV肝素结合位点与细胞之间的结合，从而允许AAV进入上清液，即培养基 。 因此，与使用细胞裂解步骤的方法产生的AAV相比，本发明的方法提供了含有高产率的AAV的上清液，其具有比细胞膜和细胞内物质更高的纯度。