Production of a sialylated N-linked glycoprotein in insect cells.
Under High Aspect Ratio Vessel (HARV) bioreactor culture conditions designed to simulate the microgravity of orbital space flight, insect tissue culture cells infected with a baculovirus expression vector produced a human glycoprotein with tri- and tetra-antennary complex N-linked oligosaccharides containing terminal sialic acid residues. The oligosaccharide structures were similar to those produced in human placental cells. Insect cells cultured in T-flasks only performed incomplete oligosaccharide processing. The mechanism of HARV-mediated changes in the eukaryotic N-linked glycosylation pathway was investigated and could be mimicked under T-flask growth conditions with the addition of N-acetylmannosamine to the culture medium. The significance of these investigations is discussed with respect to the production of recombinant therapeutic glycoproteins, insect physiology, and orbital space flight.
Joshi
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Boyce Thompson Institute at Cornell University and School of Chemical Engineering, Cornell University, 120 Olin Hall, Ithaca, New York 14853, USA. Lokesh.Joshi@asu.edu
Shuler
M L
ML
Wood
H A
HA
D000818:Animals; D019149:Bioreactors; D002851:Chromatography, High Pressure Liquid; D006023:Glycoproteins / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics; D006595:Hexosamines / Q000032:analysis; D006801:Humans; D007313:Insecta / Q000166:cytology* / Q000378:metabolism; D019158:N-Acetylneuraminic Acid / Q000032:analysis*; D014161:Transduction, Genetic; D014893:Weightlessness
D006023:Glycoproteins; D006595:Hexosamines; D019158:N-Acetylneuraminic Acid; C002022:N-acetylmannosamine