02290nas a2200229   4500008004100000022001400041245008800055210006900143260000800212300001000220490000700230520157600237653001101813653002001824653001801844653001101862653002401873653001701897100001501914700001501929856011601944       2019                        eng d  a0966-084400aNickel and cobalt affect galactosylation of recombinant IgG expressed in CHO cells.0 aNickel and cobalt affect galactosylation of recombinant IgG expr  cOCT  a11-190 v323 a<p>Glycosylation is an important product quality attribute of antibody biopharmaceuticals. It involves enzymatic addition of oligosaccharides on proteins by sequential action of glycosyltransferases and glycosidases in the endoplasmic reticulum and golgi. Some of these enzymes like galactosyltransferase and N-acetylglucosaminyltransferase-I require trace metal cofactors. Variations in trace metal availability during production can thus affect glycosylation of recombinant glycoproteins such as monoclonal antibodies. Variability in trace metal concentrations can be introduced at multiple stages during production such as due to impurities in raw materials for culture medium and leachables from bioreactors. Knowledge of the effect of various trace metals on glycosylation can help in root-cause analysis of unintended variability in glycosylation. In this study, we investigated the effect of nickel and cobalt on glycosylation of recombinant IgG expressed in Chinese hamster ovary cells. Nickel concentrations below 500 µM did not affect glycosylation, but above 500 µM it significantly decreases galactosylation of IgG. Cobalt at 50 µM concentration causes slight increase in G1F glycans (mono galactosylated) as previously reported. However, higher concentrations result in a small increase in G0F (non galactosylated) glycans. This effect of nickel and cobalt on galactosylation of recombinant IgG can be reversed by supplementation of uridine and galactose which are precursors to UDP-Galactose, a substrate for the enzymatic galactosylation reaction.</p>
10acobalt10aGalactosylation10aglycosylation10aNickel10aProcess variability10aTrace metals1 aPrabhu, A.1 aGadgil, M.  uhttps://library.ncl.res.in/content/nickel-and-cobalt-affect-galactosylation-recombinant-igg-expressed-cho-cells