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.