在 CHO 培养物中，增强的促红细胞生成素异质性是由蛋白水解降解引起的，可以通过高谷氨酰胺水平消除。
Enhanced erythropoietin heterogeneity in a CHO culture is caused by proteolytic degradation and can be eliminated by a high glutamine level.
机构信息
Department of Microbiology, University of Manitoba, Winnipeg, Manitoba, R3T 2N2, Canada.
出版信息
Cytotechnology. 2000 Oct;34(1-2):83-99. doi: 10.1023/A:1008137712611.
The molecular heterogeneity of recombinant humanerythropoietin (EPO) increased during the course of abatch culture of transfected Chinese hamster ovary(CHO) cells grown in serum-free medium. This wasshown by both an increased molecular weight and pIrange of the isolated EPO at the end of the culture. However, analysis of the N-glycan structures of themolecule by fluorophore-assisted carbohydrateelectrophoresis (FACE) and HPLC anion exchangechromatography indicated a consistent pattern ofglycosylation. Seven glycoforms were identified, thepredominant structure being a fully sialylatedtetra-antennary glycan. The degree of sialylationwas maintained throughout the culture. Analysis ofthe secreted EPO indicated a time-dependent increasein the molecular weight band width of the peptideconsistent with proteolytic degradation. A highglutamine concentration (16-20 mM) in the culturedecreased the apparent degradation of the EPO.
在无血清培养基中培养的转染中国仓鼠卵巢（CHO）细胞的批培养过程中，重组人红细胞生成素（EPO）的分子异质性增加。这一点在培养结束时分离出的 EPO 的分子量和等电点范围增加上均有体现。然而，通过荧光辅助碳水化合物电泳（FACE）和高效阴离子交换色谱分析该分子的 N-聚糖结构表明，糖基化模式一致。鉴定出 7 种糖型，主要结构为完全唾液酸化的四天线聚糖。在整个培养过程中，唾液酸化程度得以维持。对分泌的 EPO 的分析表明，与蛋白水解降解一致，肽的分子量带宽度随时间呈依赖性增加。培养物中高浓度的谷氨酰胺（16-20 mM）可降低 EPO 的明显降解。
Zotero 插件