02344nas a2200241   4500008004100000022001400041245016400055210006900219260007100288300001200359490000700371520145300378653000801831653001301839653001401852653001201866653001801878653002401896653001401920100001801934700001901952856013101971       2015                        eng d  a8756-793800aManganese increases high mannose glycoform on monoclonal antibody expressed in cho when glucose is absent or limiting: implications for use of alternate sugars0 aManganese increases high mannose glycoform on monoclonal antibod  a111 RIVER ST, HOBOKEN 07030-5774, NJ USAbWILEY-BLACKWELLcMAR-APR  a460-4670 v313 a<p>Alternate sugars such as galactose and fructose are metabolized at a slower rate than glucose and result in lower accumulation of lactate. While low lactate accumulation is desirable, we report that complete substitution of glucose with these sugars results in an increase in M5 high mannose glycans. Surprisingly, this increase is much higher when the culture is supplemented with manganese: for example, when cells are cultured with galactose, M5 high mannose glycan content increased from 5% at 1 nM Mn2+ in the basal medium to 32% with 16 mu M Mn2+ supplementation. When galactose is supplemented with glucose maintained at low concentrations, a small reduction in high mannose glycans is seen. In control cultures with glucose, the high mannose content was however &lt;2% in this range of Mn2+ concentration. By varying Mn2+ and glucose supplementation levels, with or without galactose, we systematically demonstrate that Mn2+ concentration and glucose availability, together, significantly affect the high mannose glycan content. To our knowledge, this is the first report that shows that the effect of Mn2+ on high mannose glycan content depends on glucose availability. At each Mn2+ supplementation level evaluated, galactosylation percentages were highest for cultures where galactose was supplemented with glucose at non-limiting concentration. (c) 2014 American Institute of Chemical Engineers Biotechnol. Prog., 31:460-467, 2015</p>10aCHO10afructose10agalactose10aglucose10aglycosylation10ahigh-mannose glycan10amanganese1 aSurve, Tanaya1 aGadgil, Mugdha  uhttps://library.ncl.res.in/content/manganese-increases-high-mannose-glycoform-monoclonal-antibody-expressed-cho-when-glucose-002503nas a2200217   4500008004100000022001200041245008300053210006900136260000800205300000900213520182000222653001402042653001802056653001402074653001702088653000902105100001802114700002202132700001902154856011202173       2018                        eng d  a175759800aZinc supplementation decreases galactosylation of recombinant IgG in CHO cells0 aZinc supplementation decreases galactosylation of recombinant Ig  cMAY  a1-113 a<p>Trace element composition of culture medium can be altered to modulate glycoform of recombinant glycoproteins. In this study, we show that Zn2+ supplementation at or above 100 μM decreases galactosylation of recombinant IgG expressed in Chinese Hamster Ovary cells. This decrease in galactosylation is not due to reduced galactosyltransferase expression. This effect persists upon supplementation of galactose and uridine to the culture, indicating that it may not be due to reduced UDP-Gal availability. Measurements of galactosyltransferase activity in the cell lysate show that activity decreases with increasing Zn2+/Mn2+ ratio. This suggests that one possible explanation of the effect of Zn2+ may be reduced intracellular galactosyltransferase activity due to increase in Zn2+/Mn2+ ratio. Consistent with this, the decrease in galactosylation of IgG could be reversed by supplementation of Mn2+ (a cofactor of galactosyltransferase) which increases intracellular Mn2+ content. Measurement of total intracellular Zn2+ content, however, indicates no significant upregulation of total intracellular Zn2+ content and no significant downregulation of intracellular Mn2+ content with Zn2+ supplementation. One possible explanation could be that cellular detoxification response to higher extracellular Zn2+ concentration might lead to changes in intracellular distribution of Mn2+. In this case, Zn2+ supplementation would be expected to interfere with other known effects of Mn2+. Indeed, the previously reported increase in high mannose glycans upon Mn2+ supplementation in the absence of glucose is reversed by Zn2+ supplementation. This study also suggests the use of Mn2+ supplementation as a strategy to overcome the effect of lot-to-lot variability in trace element concentrations on galactosylation.</p>10aCHO cells10aglycosylation10amanganese10aTrace metals10azinc1 aPrabhu, Anuja1 aGadre, Ramchandra1 aGadgil, Mugdha  uhttps://library.ncl.res.in/content/zinc-supplementation-decreases-galactosylation-recombinant-igg-cho-cells