公开(公告)号：US20160040124A1

公开(公告)日：2016-02-11

申请号：US14775154

申请日：2014-03-13

申请人： Carrie L. BALLIET ,  James M. WAGNER ,  Shyamsundar SUBRAMANIAN ,  David PAJEROWSKI ,  Danilo CASIMIRO ,  Jessica FLYNN ,  Jennifer GALLI ,  Patrick MCHUGH ,  Dawn HALL ,  Christopher DANIELS ,  Jennifer GIRARD ,  MERCK SHARP & DOHME CORP.

发明人： James M. Wagner ,  Shyamsundar Subramanian ,  David Pajerowski ,  Danilo Casimiro ,  John Balliet (DECEASED) ,  Jessica Flynn ,  Jennifer Galli ,  Patrick McHugh ,  Dawn Hall ,  Christopher Daniels ,  Jennifer Girard

IPC分类号： C12N5/07 ,  C12P21/00 ,  C07K14/005 ,  C12N7/00

CPC分类号： C12N5/0601 ,  C07K14/005 ,  C12N7/00 ,  C12N2500/60 ,  C12N2511/00 ,  C12N2770/36151 ,  C12P21/00

摘要： The present invention relates to the use of increased culture pH, relative to standard insect cell culture conditions, during baculovirus infection of lepidopteran insect cells to enable production of recombinant chikungunya (CHIKV) virus like particles (VLPs). The invention further relates to adapted insect cell lines derived from insect cells such as Sf21, which can grow robustly at elevated culture pH, the use of said cell lines to recombinantly produce pH sensitive proteins in the correct conformation and increase expression of recombinant proteins relative to standard insect cell lines. In some embodiments of the invention, the cells are useful for recombinant production of CHIKV VLPs. The invention also relates to a method for the production of a pH-adapted lepidopteran insect cell line. In some embodiments of said method, the cell line is produced and/or maintained in reduced phosphate serum-free insect cell media.

摘要翻译： 本发明涉及在鳞翅目昆虫细胞杆状病毒感染期间相对于标准昆虫细胞培养条件增加培养物pH的用途，以使得能够生产重组基孔肯雅（CHIKV）病毒样颗粒（VLP）。 本发明还涉及衍生自昆虫细胞如Sf21的适应性昆虫细胞系，其可以在升高的培养pH下强壮生长，使用所述细胞系以正确构象重组产生pH敏感蛋白并增加重组蛋白相对于 标准昆虫细胞系。 在本发明的一些实施方案中，所述细胞可用于重组生产CHIKV VLP。 本发明还涉及一种用于生产pH适应的鳞翅目昆虫细胞系的方法。 在所述方法的一些实施方案中，产生和/或维持细胞系在减少的无磷酸血清无血清昆虫细胞培养基中。