Zinc supplementation decreases galactosylation of recombinant IgG in CHO cells

TitleZinc supplementation decreases galactosylation of recombinant IgG in CHO cells
Publication TypeJournal Article
Year of Publication2018
AuthorsPrabhu, A
Secondary AuthorsGadre, R
Tertiary AuthorsGadgil, M
JournalApplied Microbiology and Biotechnology
Pagination1-11
Date PublishedMAY
Type of ArticleJournal Article
ISSN1757598
KeywordsCHO cells, glycosylation, manganese, Trace metals, zinc
Abstract

<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>

DOI10.1007/s00253-018-9064-8
Type of Journal (Indian or Foreign)

Foreign

Impact Factor (IF)

3.420

Divison category: 
Chemical Engineering & Process Development