The effect of dissolved oxygen (DO) concentration on the glycosylation of recombinant protein produced by the insect cell-baculovirus expression system. |
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Authors: | Fuming Zhang Mark A Saarinen Laura J Itle Sybil C Lang David W Murhammer Robert J Linhardt |
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Institution: | Department of Chemical and Biochemical Engineering, University of Iowa, Iowa City, Iowa 52242, USA. |
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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. |
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Keywords: | human secreted alkaline phosphatase (SEAP) baculovirus expression vector system (BEVS) insect cells protein glycosylation dissolved oxygen (DO) concentration |
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