Kang Sohye, Zhang Zhongqi, Richardson Jason, Shah Bhavana, Gupta Shivani, Huang Chung-Jr, Qiu Jinshu, Le Nicole, Lin Henry, Bondarenko Pavel V
Materials and Systems Analytics, Amgen Inc., One Amgen Center Drive, Thousand Oaks, CA 91320, USA.
Materials and Systems Analytics, Amgen Inc., One Amgen Center Drive, Thousand Oaks, CA 91320, USA.
J Biotechnol. 2015 Jun 10;203:22-31. doi: 10.1016/j.jbiotec.2015.03.002. Epub 2015 Mar 19.
High mannose (HM) glycan levels on secreted monoclonal antibodies can be influenced by external factors, including osmolality and copper deficiency, and by intrinsic factors determined by different cell lines. In order to identify the metabolic markers associated with HM glycan levels, metabolomics analysis was performed to assess the changes in the extracellular metabolites of recombinant cell lines at different time points during fed-batch production process. Ornithine was identified as the common metabolic marker influenced by both external and intrinsic factors when eight different medium conditions and eight different cell lines exhibiting different levels of HM were compared. A strong correlation was also observed between HM and mRNA expression levels of arginase 1, an enzyme that catalyzes the conversion of arginine to ornithine. The results from functional validation study showed that the supplementation of ornithine to the culture medium leads to an increased level of HM, while reduced concentration of spermine, a downstream product of ornithine metabolism, leads to a decreased level of HM. Additional metabolic markers correlating with HM glycan levels were identified from eight-cell line comparison analysis. A common feature shared by these identified markers is their previously described roles as contributors of cellular redox regulation.
分泌型单克隆抗体上的高甘露糖（HM）聚糖水平会受到外部因素（包括渗透压和铜缺乏）以及不同细胞系所决定的内在因素的影响。为了识别与HM聚糖水平相关的代谢标志物，进行了代谢组学分析，以评估分批补料生产过程中不同时间点重组细胞系细胞外代谢物的变化。当比较八种不同的培养基条件和八种表现出不同HM水平的不同细胞系时，鸟氨酸被确定为受外部和内在因素共同影响的常见代谢标志物。在HM与精氨酸酶1的mRNA表达水平之间也观察到了强烈的相关性，精氨酸酶1是一种催化精氨酸转化为鸟氨酸的酶。功能验证研究的结果表明，向培养基中添加鸟氨酸会导致HM水平升高，而鸟氨酸代谢的下游产物精胺浓度降低会导致HM水平降低。从八细胞系比较分析中鉴定出了与HM聚糖水平相关的其他代谢标志物。这些鉴定出的标志物的一个共同特征是它们先前被描述为细胞氧化还原调节的贡献者。
