title: Engineering N-Glycosylation Pathway in Insect Cells: Suppression of β-N-Acetylglucosaminidase and Expression of β-1,4-Galactosyltransferase.
abstract: Most insect cells have a simple N-glycosylation process and consequently paucimannosidic or simple core glycans predominate. It has been proposed that β-N-acetylglucosaminidase (GlcNAcase), a hexosaminidase in the Golgi membrane which removes a terminal N-acetylglucosamine (GlcNAc), might contribute to simple N-glycosylation profile in several insect cells including Drosophila S2. Here, we describe GlcNAcase suppression strategy using RNA interference (RNAi) to avoid the formation of paucimannosidic glycans in insect S2 cells. In addition, we describe coexpression of β(1,4)-galactosyltransferase (GalT) as a strategy to improve N-glycosylation pattern and enable recombinant therapeutic proteins to be produced in S2 cells with more complex N-glycans.
authors.affiliation: Department of Chemical Engineering, Pohang University of Science and Technology, Pohang, Korea.
authors.forename: Yeon Kyu
authors.initials: YK
authors.lastname: Kim
authors.forename: Hyung Joon
authors.initials: HJ
authors.lastname: Cha
mesh_terms: D000118:Acetylglucosaminidase / Q000235:genetics*; D000818:Animals; D006031:Glycosylation; D007313:Insecta / Q000235:genetics*; D009237:N-Acetyllactosamine Synthase / Q000235:genetics*; D011134:Polysaccharides / Q000235:genetics; D034622:RNA Interference / Q000502:physiology; D011994:Recombinant Proteins / Q000235:genetics; D001619:beta-N-Acetylhexosaminidases / Q000235:genetics*
chemical_list: D011134:Polysaccharides; D011994:Recombinant Proteins; D009237:N-Acetyllactosamine Synthase; D000118:Acetylglucosaminidase; D001619:beta-N-Acetylhexosaminidases