金属离子对CHO细胞抗体表达及抗体电荷分布的影响

doi:10.13560/j.cnki.biotech.bull.1985.2016.08.034

摘要/Abstract

摘要： 旨在深入认识金属离子在中国仓鼠卵巢（CHO）细胞培养生产单克隆抗体过程中所发挥的作用。综合考察了不同铜离子和锌离子浓度下CHO细胞的生长、抗体的表达以及抗体的电荷分布情况。结果显示,一方面,当培养基中铜离子浓度为120 nmol/L、锌离子浓度为50 µmol/L时,最有利于CHO细胞的生长和抗体的表达。过高或过低的铜离子和锌离子均对细胞的生长和活性的维持产生了不利影响。另一方面,作为碱性羧肽酶抑制剂和辅因子的铜离子和锌离子对抗体的电荷分布有着重要的影响。当培养基中铜离子浓度为50 nmol/L、锌离子浓度为50 µmol/L时,最有利于减少抗体的电荷变体,提高主峰含量。过高或过低的铜离子和锌离子均导致了电荷变体的增加,主峰含量的降低。且两者的浓度和比例影响了抗体C末端赖氨酸的酶切过程,进而影响了碱性变体的含量。此外,对铜、锌离子的操作空间进行了初步的研究。金属离子在CHO细胞培养过程中对细胞的生长、抗体的表达以及抗体的电荷分布等方面都发挥着重要的作用。

关键词: 动物细胞培养, 电荷分布, 金属离子, 单克隆抗体

Abstract: The objective of this work is to get insights into the role of metal ions in the production of the monoclonal antibody（mAb）during Chinese hamster ovary（CHO）cell cultures. Cell growth,antibody production and antibody charge distribution at different concentrations of copper and zinc ions during cell cultures were comprehensively investigated. It was found that 120 nmol/L copper ion and 50 µmol/L zinc ion were the optimal concentrations for cell growth and antibody production. Excessively low or high levels of copper and zinc ions brought negative impacts on cell growth and viability maintenance. Additionally,copper ion and zinc ion,as the inhibitor and cofactor of basic carboxypeptidase respectively,showed considerable effects on antibody charge distribution. The medium containing 50 nmol/L copper ion and 50 μmol/L zinc ion was the most conducive to the reduction of charge variants and the increase of main species. Both excessively low or high concentrations of copper and zinc ions led to the increase of the antibody charge variants and the reduction of main species. The concentrations and ratios of copper and zinc ions altered the enzymatic digestion at antibody C-terminal lysine,subsequently affected the content of the basic variant. Additionally,the working space of copper and zinc ions for process control of antibody charge variants was preliminarily discussed. In conclusion,the metal ions play important roles in cell growth,mAb production and mAb charge distribution during CHO cell cultures.

Key words: animal cell culture, charge distribution, metal ions, monoclonal antibody

张鑫涛,唐红萍,赵亮,范里,刘旭平,缪仕伟,谭文松. 金属离子对CHO细胞抗体表达及抗体电荷分布的影响[J]. 生物技术通报, 2016, 32(8): 233-241.

ZHANG Xin-tao ,TANG Hong-ping ,ZHAO Liang ,FAN Li, LIU Xu-ping, MIAO Shi-wei, TAN Wen-song. Effects of Metal Ions on Antibody Production and Charge Heterogeneity in CHO Cell Cultures[J]. Biotechnology Bulletin, 2016, 32(8): 233-241.

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