公开(公告)号：US5994134A

公开(公告)日：1999-11-30

申请号：US73076

申请日：1998-05-04

申请人： Daniel D. Giroux ,  Ann M. Goudreau ,  Muralidhara Ramachandra ,  Paul W. Shabram

发明人： Daniel D. Giroux ,  Ann M. Goudreau ,  Muralidhara Ramachandra ,  Paul W. Shabram

IPC分类号： C12N15/09 ,  C12N5/10 ,  C12N7/00 ,  C12N7/01

CPC分类号： C12N7/00 ,  A61K48/0091 ,  C12N15/86 ,  C12N2531/00 ,  C12N2710/10043 ,  C12N2710/10343 ,  Y10S435/948

摘要： The present invention is directed to a method of producing recombinant viral vectors at high titers incorporating a variety of important advancements over the art. The method of the present invention incorporates multiple features which provide enhanced production of viruses, particularly those viruses encoding exogenous transgenes. The specifically illustrated method describes a method for the high titer serum-free media production of recombinant replication defective adenoviruses containing an exogenous transgene. The invention provides methods of preparing microcarriers, methods for seeding bioreactors at high cell density, increasing the infectivity of the producer cells to the virus, methods to increase product yield through synchronization of the cell cycle of the producer cells, and methods to minimize the deleterious effects of exogenous transgenes. The invention further provides producer cells prepared by the process of the invention. The invention further provides viruses produced by the process.

摘要翻译： 本发明涉及以高滴度生产重组病毒载体的方法，其结合了本领域的各种重要进展。 本发明的方法结合了多个特征，其提供增强的病毒生产，特别是编码外源转基因的那些病毒。 具体说明的方法描述了含有外源转基因的重组复制缺陷型腺病毒的高滴度无血清培养基的制备方法。 本发明提供了制备微载体的方法，用于以高细胞密度接种生物反应器的方法，增加生产细胞对病毒的感染性，通过生产细胞的细胞周期同步来提高产量的方法，以及使有害细菌最小化的方法 外源转基因的作用。 本发明还提供了通过本发明的方法制备的生产细胞。 本发明还提供了通过该方法产生的病毒。