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The effect of dissolved oxygen (DO) concentration on the glycosylation of recombinant protein produced by the insect cell-baculovirus expression system.
Authors:Fuming Zhang  Mark A Saarinen  Laura J Itle  Sybil C Lang  David W Murhammer  Robert J Linhardt
Institution:Department of Chemical and Biochemical Engineering, University of Iowa, Iowa City, Iowa 52242, USA.
Abstract:The effect of dissolved oxygen concentration on human secreted alkaline phosphatase (SEAP) glycosylation by the insect cell-baculovirus expression system was investigated in a well-controlled bioreactor. Oligomannose-type N-linked glycans (i.e., Man2 to Man6 and Man3F) were present in SEAP produced by Spodoptera frusiperda Sf-9 (Sf-9) and Trichoplusia ni BTI-Tn-5B1-4 (Tn-5B1-4) insect cell lines. The relative amounts of the most highly processed glycans (i.e., Man3F and Man2 in the SEAP from Sf-9 and Tn-5B1-4 cells, respectively) were significantly higher at 50% of air saturation than at either 10% or 190% of air saturation. That is, glycan processing was inhibited at both low and high dissolved oxygen concentrations.
Keywords:human secreted alkaline phosphatase (SEAP)  baculovirus expression vector system (BEVS)  insect cells  protein glycosylation  dissolved oxygen (DO) concentration
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