The effect of cell concentration on alpha 2,3-sialyltransferase activity in attachment culture of a human erythropoietin-producing Chinese hamster ovary cell line |
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Authors: | Hyoung Jin Kim Hong-Jin Kim |
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Institution: | (1) Department of Chemical and Biomolecular Engineering, Johns Hopkins University, 3400 North Charles Street – Baltimore, Maryland, 21218, USA; |
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Abstract: | Terminal sialylation of therapeutic glycoprotein is important for biological activity and in vivo stability. The enzyme α2,3-sialyltransferase is the key enzyme that links sialic acids to the termini of glycans in the Chinese
hamster ovary (CHO) cell line. Terminal sialylation is affected by numerous factors, but the elements that regulate α2,3-sialyltransferase
are not known. We investigated the relationship between α2,3-sialyltransferase activity, ammonium concentration, and cell
attachment area-based cell concentration in a recombinant human erythropoietin (rhEPO)-producing CHO cell line. We found that
ammonium in the culture medium had almost no effect on α2,3-sialyltransferase activity, but that the activity was affected
by cell attachment area-based cell concentration; α2,3-sialyltransferase activity and terminal sialylation of rhEPO decreased
with increasing the cell concentration. These results demonstrate that the cell attachment area-based cell concentration is
an important factor that affects 2,3-sialyltransferase activity and terminal sialylation of CHO cells. |
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