Effect of Zn2+on IgG2 Expression and Disulfide Bond Isoforms Distribution in CHO Cells
Objective:To investigate the effects of Zn2+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 200 μmol/L,Zn2+has certain negative effects on CHO cell growth,viability maintenance and IgG2 antibody expression,and is negatively correlated with Zn2+concentration.The addition of Zn2+can significantly increase the proportion of IgG2-B isoforms in IgG2 disulfide bond isomer,and down-regulate the proportion of IgG2-A and IgG2-A/B isoforms.Among them,100 μmol/L Zn2+has the most obvious effect,and the proportion of IgG2-B increase by nearly 10 percentage points.Conclusion:Zn2+can inhibit the growth of CHO cells and the expression of IgG2,and Zn2+is also important for the regulation of disulfide isoforms.
CHO cellimmunoglobulin G2Zn2+disulfide bond isoforms
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