Effect of cysteamine on cell growth and IgG[4] production in recombinant Sp2.0 cells

ABSTRACT

The manipulation of redox potential in secretory pathway by thiol reducing agents can be a strategy to improve the production levels of disulfide-bonded proteins including recombinant antibodies. Here we have studied the influence of cysteamine on viability and the production level of IgG[4] in Sp2.0 cells. For this purpose, the recombinant Sp2.0 cells producing an anti CD33 IgG[4], were subjected to different concentrations of cysteamine. At concentrations of 2, 4 and 5 mM cysteamine, the secreted levels of IgG[4] did not change significantly. However, in concentration of 7 mM cysteamine, a significant decrease was observed in IgG[4] levels which may indicate the cytotoxicity of this compound in higher concentrations. Our results show that the cysteamine treatment reduces the cell viability in a dose-dependent manner. Also it was observed that 2 mM cysteamine had no late effect on IgG4 production level and only at day 3, this concentration of cysteamine decreased the cell viability significantly. To test whether the addition of cysteamine can affect the expression level of protein disulfide isomerase, RT-PCR analysis was carried out. The results revealed that cysteamine does not affect the PDI transcription and expression level of IgG[4] in this type of recombinant cells