![]() Flow cytometry was selected as the read-out technique for rxYFP signal given its high-throughput and statistical robustness. In this study, we validate the use of cytosolic rxYFP on two cell lines widely used in biomanufacturing processes, namely, CHO-K1 cells expressing the human granulocyte macrophage colony-stimulating factor (hGM-CSF) and HEK-293. ![]() ![]() Recently, different redox-sensitive variants of green fluorescent proteins (e.g., rxYFP, roGFP2, and rxmRuby2) have been engineered and proved suitable to detect, in a non-invasive manner, perturbations in the pool of reduced and oxidized glutathi-one, the major low molecular mass thiol in mammals. Thus, monitoring redox changes in real time and in situ is deemed essential for optimizing the production of recombinant proteins. Exogenous (i.e., nutrients, deterioration of media components, xenobiotics) and endogenous factors (i.e., metabolism, growth) may alter the redox homeo-stasis of cells. Cellular functions such as DNA replication and protein translation are influenced by changes in the intracellular redox milieu.
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