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    • 9. 发明公开
    • EXPRESSION VECTOR ELEMENT COMBINATIONS, NOVEL PRODUCTION CELL GENERATION METHODS AND THEIR USE FOR THE RECOMBINANT PRODUCTION OF POLYPEPTIDES
    • 表达载体的元素组合,生产电池生产及新方法它们在重组多肽生产
    • EP2794651A2
    • 2014-10-29
    • EP12810245.6
    • 2012-12-19
    • F.Hoffmann-La Roche AG
    • HUELSMANN, Peter MichaelKNOETGEN, Hendrik
    • C07K16/00C12N15/67
    • C12N15/85C07K16/00C07K16/2854C07K16/468C07K2317/14C12N2830/00C12N2830/36C12N2830/42C12N2840/203
    • Herein is reported that for transient transfections the use of the human elongation factor 1 alpha promoter (with Intron A) provides for an enhanced productivity (in LC-HC-SM organization), the use of the bovine growth hormone polyA signal sequence provides for an enhanced productivity compared to use of the SV40 polyA signal sequence, the addition of the HGT to the bGH PolyA signal sequence results in an increased productivity in vectors containing the hCMV promoter and the vector organization LC(3′-5′)-HC-SM results in improved expression. For stable pools it is reported that pools generated with vectors containing the hEF1α promoter show an enhanced productivity in batch analysis, clones generated with vectors containing the hEF1α promoter show a reduced number of low producing clones, and clones generated with vectors containing the hEF1α promoter show a higher stability of IgG expression. For single clones it is reported that the vector organization with downstream position of selection marker (LC-HC-SM) has a positive effect on productivity of single clones and that clones generated with vectors containing the bGH polyA signal sequence and the hGT have higher productivities.
    • 在报道没有为瞬时转染使用人延伸因子1α启动子(内含子A)的提供用于上提高生产率(在LC-HC-SM组织),使用牛生长激素多聚腺苷酸信号序列提供了用于 相比于使用SV40的增强生产力多聚A信号序列,添加HGT到的bGH polyA信号序列中的生产力提高导致在含有hCMV启动子与矢量组织LC(3'-5“)载体 - HC-SM 导致改进的表达。 对于稳定池有报道并与载体通过含有HEF1α启动子节目以提高生产率在分批分析产生池,用含有HEF1α启动子显示出低生产克隆的数量减少的矢量产生的克隆,并用载体通过含有HEF1α启动子节目产生克隆 IgG表达的更高的稳定性。 对于单克隆有报告做了与选择标记物(LC-HC-SM)的下游位置的矢量组织对单个克隆的生产率具有积极作用,并用含有所述的bGH polyA信号序列和HGT矢量生成并克隆具有更高的生产率 ,