首页> 外文会议>Cell culture engineering conference >GENOTYPE OF CHO HOST CELL LINE HAS HIGHER IMPACT ON MAB PRODUCTION AND QUALITY THAN PROCESS STRATEGY OR CELL CULTURE MEDIUM
【24h】

GENOTYPE OF CHO HOST CELL LINE HAS HIGHER IMPACT ON MAB PRODUCTION AND QUALITY THAN PROCESS STRATEGY OR CELL CULTURE MEDIUM

机译:CHO宿主细胞系的基因型比过程策略或细胞培养培养基对单克隆抗体生产和质量的影响更大

获取原文

摘要

Chinese hamster ovary (CHO) cells comprise a variety of lineages, including CHO-DXB11, CHO-K1, CHO-DG44 and CHO-S. Despite the fact that CHO cell lines share a common ancestor, extensive mutagenesis and clonal selection have resulted in substantial genetic heterogeneity among them. Data from sequencing shows that different genes are lacking from individual CHO cell lines and that each cell line harbors a unique set of mutations that are relevant to the bioprocess. However, literature outlining how the observed genetic differences affect CHO cell performance during bioprocess operations remains scarce. In this study, we examined host cell-specific differences among three widely used CHO cell lines (CHO-K1, CHO-S and CHO-DG44) and recombinantly expressed the same monoclonal antibody (mAb) in an isogenic format in all cell lines by using bacterial artificial chromosomes (BACs) as transfer vector. Cell-specific growth, product formation and heavy and light chain mRNA levels were studied in batch, fed-batch and perfusion cultures. Furthermore, two different cell culture media were investigated. Product quality was studied through glycoprofiling, and the thermal denaturation was analyzed using differential scanning calorimetry (DSC). We found CHO cell line-specific preferences for mAb production or biomass synthesis that were determined by the host cell line rather than product-specific mRNA levels. Additionally, quality attributes of the expressed mAb were influenced by the host cell line and medium used.
机译:中国仓鼠卵巢(CHO)细胞包含多种谱系,包括CHO-DXB11,CHO-K1,CHO-DG44和CHO-S。尽管CHO细胞系具有共同的祖先这一事实,但广泛的诱变和克隆选择已导致它们之间存在实质性的遗传异质性。测序数据显示单个CHO细胞株缺少不同的基因,并且每个细胞株都具有与生物过程相关的独特突变集。但是,概述在生物过程中观察到的遗传差异如何影响CHO细胞性能的文献仍然很少。在这项研究中,我们检查了三种广泛使用的CHO细胞系(CHO-K1,CHO-S和CHO-DG44)之间的宿主细胞特异性差异,并通过所有细胞系以等基因形式重组表达了相同的单克隆抗体(mAb),使用细菌人工染色体(BAC)作为转移载体。分批,补料分批和灌注培养研究细胞特异性生长,产物形成以及重链和轻链mRNA水平。此外,研究了两种不同的细胞培养基。通过糖谱分析研究产品质量,并使用差示扫描量热法(DSC)分析热变性。我们发现mAb生产或生物质合成的CHO细胞系特异性偏好是由宿主细胞系而非产物特异性mRNA水平决定的。另外,表达的mAb的质量属性受宿主细胞系和所用培养基的影响。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱:kefu@zhangqiaokeyan.com

京公网安备:11010802029741号 ICP备案号:京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有
  • 客服微信

  • 服务号