锌离子对CHO细胞IgG2抗体表达及二硫键异构体分布的影响  

Effect of Zn^(2+)on IgG2 Expression and Disulfide Bond Isoforms Distribution in CHO Cells

作　　者：刘学明 蒋仁材 韦梦娟 刘唯 胡婷婷 郭兴东 曹春来 LIU Xueming;JIANG Rencai;WEI Mengjuan;LIU Wei;HU Tingting;GUO Xingdong;CAO Chunlai(Federal Biotechnology(Zhuhai Hengqin)Co.,Ltd.,Zhuhai 519060,China;Zhuhai Federal Biomedical Co.,Ltd.,Zhuhai 519040,China)

机构地区：[1]联邦生物科技(珠海横琴)有限公司,广东珠海519060 [2]珠海联邦生物医药有限公司,广东珠海519040

出　　处：《生物化工》2024年第3期14-19,34,共7页Biological Chemical Engineering

摘　　要：目的:研究Zn^(2+)对中国仓鼠卵巢细胞(CHO细胞)生长、IgG2表达及其二硫键异构体分布的影响。方法:补料分批培养外分泌IgG2的CHO细胞株中,分别添加不同浓度的葡萄糖酸锌,每天监测细胞密度以及活率;培养14 d后离心除细胞得上清液,通过生化分析仪测定上清液中IgG2含量,之后应用蛋白A磁珠亲和得到IgG2纯化样品,非还原十二烷基硫酸钠毛细管电泳(nrCE-SDS)、阳离子交换高效液相色谱(CEX-HPLC)和反相高效液相色谱(RP-HPLC)检测IgG2二硫键异构体分布。结果:25~200μmol/L浓度条件下,Zn^(2+)对于CHO细胞生长,活率维持以及IgG2抗体的表达均有一定的负面影响,并与Zn^(2+)浓度具有负相关性;而Zn^(2+)的添加能够显著提高IgG2二硫键异构体中IgG2-B亚型的比例,下调IgG2-A和IgG2-A/B两种亚型比例。其中100μmol/L Zn^(2+)效果最明显,IgG2-B比例提高近10个百分点。结论:Zn^(2+)能够抑制CHO细胞生长以及IgG2表达,同时Zn^(2+)对二硫键异构体的调控至关重要。Objective:To investigate the effects of Zn^(2+)on the growth,IgG2 expression and distribution of disulfide isoforms in Chinese Hamster Ovary cell(CHO cell).Methods:CHO cell lines exocrine IgG2 are cultured with different concentrations of zinc gluconate,and the cell density and viability are monitored daily.After 14 days of culture,the cells are centrifuged to obtain the supernatant,and the IgG2 content in the supernatant is determined by biochemical analyzer.After that,the purified IgG2 samples are obtained by using Protein A MagBeads affinity.The distribution of IgG2 disulfide isoforms is detected by non-reducing sodium dodecyl sulfate capillary electrophoresis(nrCE-SDS),cation-exchange high-performance liquid chromatography(CEX-HPLC),and reversed-phase high-performance liquid chromatography(RP-HPLC).Results:In the range of 25 to 200μmol/L,Zn^(2+)has certain negative effects on CHO cell growth,viability maintenance and IgG2 antibody expression,and is negatively correlated with Zn^(2+)concentration.The addition of Zn^(2+)can significantly increase the proportion of IgG2-B isoforms in IgG2 disulfide bond isomer,and downregulate the proportion of IgG2-A and IgG2-A/B isoforms.Among them,100μmol/L Zn^(2+)has the most obvious effect,and the proportion of IgG2-B increase by nearly 10 percentage points.Conclusion:Zn^(2+)can inhibit the growth of CHO cells and the expression of IgG2,and Zn^(2+)is also important for the regulation of disulfide isoforms.

关 键 词：CHO细胞 免疫球蛋白G2 锌离子 二硫键异构体 

分 类 号：Q815[生物学—生物工程]