发明名称 |
ENHANCED PRODUCTION OF RECOMBINANT PROTEINS BY TRANSIENT TRANSFECTION OF SUSPENSION-GROWING MAMMALIAN CELLS |
摘要 |
Disclosed is a new process for the production of recombinant proteins, by transient transfection of suspension-grown human embryonic kidney cells (293 cell line and its genetic variants) with an expression vector, using polyethylenimine (PEl) as a transfection reagent. In a preferred embodiment, the process uses 293E cells expressing the Epstein-Barr virus (EBV) EBNA 1 protein, in combi nation with an oriP-based episomal expression vector having an improved cytomegalovirus expression cassette comprising the CMV5 promoter. The process combines in a single step the cell growth, transfection and protein expression, is carried out without changing the culture medium, and allows to achieve high expression levels in a short period of time. The process may be carried out in a serum-free, low-protein culture medium, is easily scalable, compatible with continuous production processes, and fully adapted to high-throughput production of milligram quantities of recombinant proteins. |
申请公布号 |
WO02090533(A2) |
申请公布日期 |
2002.11.14 |
申请号 |
WO2002CA00683 |
申请日期 |
2002.05.07 |
申请人 |
NATIONAL RESEARCH COUNCIL OF CANADA;DUROCHER, YVES;PERRET, SYLVIE;PHAM, PHUONG, LAN;ST-LAURENT, GILLES;KAMEN, AMINE |
发明人 |
DUROCHER, YVES;PERRET, SYLVIE;PHAM, PHUONG, LAN;ST-LAURENT, GILLES;KAMEN, AMINE |
分类号 |
C12N5/071;C12N15/85;C12P21/02 |
主分类号 |
C12N5/071 |
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