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丁酸钠对重组中国仓鼠卵巢细胞产生的抗体的组装、电荷变体和糖基化的影响。

Effect of sodium butyrate on the assembly, charge variants, and galactosylation of antibody produced in recombinant Chinese hamster ovary cells.

作者信息

Hong Jong Kwang, Lee Sang Min, Kim Kyung-Yong, Lee Gyun Min

机构信息

Department of Biological Sciences, KAIST, 335 Gwahak-ro, Yuseong-gu, Daejeon, 305-701, South Korea,

出版信息

Appl Microbiol Biotechnol. 2014 Jun;98(12):5417-25. doi: 10.1007/s00253-014-5596-8. Epub 2014 Feb 21.

Abstract

Sodium butyrate (NaBu) is known to increase the specific productivity of recombinant Chinese hamster ovary (rCHO) cells. To understand the effects of NaBu on the product quality, rCHO cells producing monoclonal antibody (Mab) were cultivated at various concentrations of NaBu (0 to 4 mM). NaBu increased correctly assembled Mab. In the absence of NaBu, the proportions of intact Mab (2H2L) and heavy chain dimer (2H) were 81 and 15 %. At 1 mM NaBu, the proportion of 2H2L increased to 93 %, whereas the proportion of 2H decreased to 2 %. No further increase in the proportion of 2H2L was obtained at a higher NaBu concentration. NaBu also affected the charge heterogeneity of Mab, which may affect the efficacy of Mab. The basic charge variants of Mabs increased with an increase in the NaBu concentration. In addition, NaBu affected the galactosylation of Mab negatively. Overall, the data obtained here show that NaBu used in rCHO cell cultures for improved Mab production affects certain quality aspects of Mab, in this case, the charge heterogeneity and galactosylation.

摘要

已知丁酸钠(NaBu)可提高重组中国仓鼠卵巢细胞(rCHO)的比生产率。为了解NaBu对产品质量的影响,将产生单克隆抗体(Mab)的rCHO细胞在不同浓度的NaBu(0至4 mM)下培养。NaBu增加了正确组装的Mab。在不存在NaBu的情况下,完整Mab(2H2L)和重链二聚体(2H)的比例分别为81%和15%。在1 mM NaBu时,2H2L的比例增加到93%,而2H的比例降至2%。在更高的NaBu浓度下,2H2L的比例没有进一步增加。NaBu还影响了Mab的电荷异质性,这可能会影响Mab的疗效。Mab的碱性电荷变体随着NaBu浓度的增加而增加。此外,NaBu对Mab的半乳糖基化有负面影响。总体而言,此处获得的数据表明,在rCHO细胞培养中用于提高Mab产量的NaBu会影响Mab的某些质量方面,在这种情况下,即电荷异质性和半乳糖基化。

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