日本語フィールド
著者：Okada T, Ihara H, Ito R, Taniguchi N, Ikeda Y.題名：Glyco-remodeling of Sf21 cell line with β-1,4-N-Acetylglucosaminyltransferase III (GnT-III)発表情報：Clinical and Translational Research on Cancer:Glycomics Applications The 2nd Seminar on Functional Glycomics for Young Investigators HGPI-Human Disease Glycomics/Proteome Initiativeキーワード：概要：抄録：The lepidopteran cell-baculovirus expression system has been widely used for the production of properly post-translationally modified protein, thus being expected to be applicable to large-scale production of mammalian glycoproteins for therapeutic and diagnostic purpose. For practical use, however, it is required to remodel N-glycosylation pathways of the host cells because pauci-mannosidic glycoform could modify functions of the expressed protein. Current knowledge indicates that specific β-N-acetylglucosaminidase is involved in trimming of the non-reducing terminal β1,2GlcNAc of core N-glycan.
To establish the cell line preferred for the production of glycoproteins with mammalian type N-glycans, we transfected Sf21 cells with rat GnT-III cDNA. GnT-III catalyzes the introduction of the bisecting GlcNAc into the N-glycan core, and thereby provides considerable protection to the β-hexosaminidase-mediated cleavage of the terminal β1,2GlcNAc residues. Structural analyses of the cellular oligosaccharide and N-glycans of the human γ-glutamyltranspeptidase expressed in Sf21 cells revealed that the stably transfected cell line predominantly produced the bisected N-glycan containing β1,2GlcNAc at α1,3Man arm. Our result indicates that GnT-III-transfection is the effective approach to the production of complex N-glycan in Sf21 cell英語フィールド
Author：Okada T, Ihara H, Ito R, Taniguchi N, Ikeda Y.Title：Glyco-remodeling of Sf21 cell line with β-1,4-N-Acetylglucosaminyltransferase III (GnT-III)Announcement information:Clinical and Translational Research on Cancer:Glycomics Applications The 2nd Seminar on Functional Glycomics for Young Investigators HGPI-Human Disease Glycomics/Proteome InitiativeAn abstract：The lepidopteran cell-baculovirus expression system has been widely used for the production of properly post-translationally modified protein, thus being expected to be applicable to large-scale production of mammalian glycoproteins for therapeutic and diagnostic purpose. For practical use, however, it is required to remodel N-glycosylation pathways of the host cells because pauci-mannosidic glycoform could modify functions of the expressed protein. Current knowledge indicates that specific β-N-acetylglucosaminidase is involved in trimming of the non-reducing terminal β1,2GlcNAc of core N-glycan.
To establish the cell line preferred for the production of glycoproteins with mammalian type N-glycans, we transfected Sf21 cells with rat GnT-III cDNA. GnT-III catalyzes the introduction of the bisecting GlcNAc into the N-glycan core, and thereby provides considerable protection to the β-hexosaminidase-mediated cleavage of the terminal β1,2GlcNAc residues. Structural analyses of the cellular oligosaccharide and N-glycans of the human γ-glutamyltranspeptidase expressed in Sf21 cells revealed that the stably transfected cell line predominantly produced the bisected N-glycan containing β1,2GlcNAc at α1,3Man arm. Our result indicates that GnT-III-transfection is the effective approach to the production of complex N-glycan in Sf21 cell