公开(公告)号：EP1590451A2

公开(公告)日：2005-11-02

申请号：EP03813433.4

申请日：2003-12-12

申请人： Alphavax, Inc. ,  Smith, Jonathan F. ,  Kamrud, Kurt ,  Dryga, Sergey ,  Alterson, Harold ,  Rayner, Jon ,  Butler, Kim ,  Maughan, Maureen F.

发明人： SMITH, Jonathan, F. ,  KAMRUD, Kurt ,  DRYGA, Sergey ,  ALTERSON, Harold ,  RAYNER, Jon ,  ALTERSON, Kim ,  MAUGHAN, Maureen, F.

IPC分类号： C12N7/02 ,  C12N15/86

CPC分类号： C12N7/00 ,  A61K48/00 ,  A61K2039/5256 ,  C12N15/86 ,  C12N2770/36143 ,  C12N2770/36151 ,  C12N2770/36162

摘要： Provided herein are methods for producing alphavirus replicon particles in high yield; replicon RNAs are electroporated into permissive cells, where the cells are at a relatively high density, together with at least one helper nucleic acid providing the necessary functions for packaging. After a growth period in appropriate medium, alphavirus replicon particles are harvested from the surfaces of the cells in which they were produced using a salt wash in which the salt concentration is from about 0.2 to about 5 M sodium chloride, calcium chloride, magnesium chloride, potassium chloride, ammonium acetate, ammonium bicarbonate, among others. After dilution, if necessary, the particles can be purified by a suitable chromatographic technique.

公开(公告)号：EP1590451B1

公开(公告)日：2015-09-16

申请号：EP03813433.4

申请日：2003-12-12

申请人： Alphavax, Inc.

发明人： SMITH, Jonathan, F. ,  KAMRUD, Kurt ,  DRYGA, Sergey ,  ALTERSON, Harold ,  RAYNER, Jon ,  ALTERSON, Kim ,  MAUGHAN, Maureen, F.

IPC分类号： C12N7/02 ,  C12N15/86

CPC分类号： C12N7/00 ,  A61K48/00 ,  A61K2039/5256 ,  C12N15/86 ,  C12N2770/36143 ,  C12N2770/36151 ,  C12N2770/36162

公开(公告)号：EP3246399A1

公开(公告)日：2017-11-22

申请号：EP17163152.6

申请日：2003-12-12

申请人： Alphavax, Inc.

发明人： SMITH, Jonathan, F. ,  KAMRUD, Kurt ,  DRYGA, Sergey ,  ALTERSON, Harold ,  RAYNER, Jon ,  ALTERSON, Kim ,  MAUGHAN, Maureen, F.

IPC分类号： C12N7/02 ,  C12N15/86

CPC分类号： C12N7/00 ,  A61K48/00 ,  A61K2039/5256 ,  C12N15/86 ,  C12N2770/36143 ,  C12N2770/36151 ,  C12N2770/36162

摘要： Provided herein are methods for producing alphavirus replicon particles in high yield; replicon RNAs are electroporated into permissive cells, where the cells are at a relatively high density, together with at least one helper nucleic acid providing the necessary functions for packaging. After a growth period in appropriate medium, alphavirus replicon particles are harvested from the surfaces of the cells in which they were produced using a salt wash in which the salt concentration is from about 0.2 to about 5 M sodium chloride, calcium chloride, magnesium chloride, potassium chloride, ammonium acetate, ammonium bicarbonate, among others. After dilution, if necessary, the particles can be purified by a suitable chromatographic technique.

摘要翻译： 本文提供了高产量生产α病毒复制子颗粒的方法; 将复制子RNA电穿孔入允许细胞中，其中细胞处于相对高的密度，连同至少一种提供包装必需功能的辅助核酸一起。 在合适的培养基中生长一段时间后，从产生它们的细胞表面收获α病毒复制子颗粒，使用盐洗涤，其中盐浓度为约0.2至约5M氯化钠，氯化钙，氯化镁， 氯化钾，乙酸铵，碳酸氢铵等。 稀释后，如果需要，可以通过合适的色谱技术来纯化颗粒。