title: Producing recombinant therapeutic glycoproteins with enhanced sialylation using CHO-gmt4 glycosylation mutant cells.
abstract: Recombinant glycoprotein drugs require proper glycosylation for optimal therapeutic efficacy. Glycoprotein therapeutics are rapidly removed from circulation and have reduced efficacy if they are poorly sialylated. Ricinus communis agglutinin-I (RCA-I) was found highly toxic to wild-type CHO-K1 cells and all the mutants that survived RCA-I treatment contained a dysfunctional N-acetylglucosaminyltransferase I (GnT I) gene. These mutants are named CHO-gmt4 cells. Interestingly, upon restoration of GnT I, the sialylation of a model glycoprotein, erythropoietin, produced in CHO-gmt4 cells was shown to be superior to that produced in wild-type CHO-K1 cells. This addendum summarizes the applicability of this cell line, from transient to stable expression of the recombinant protein, and from a lab scale to an industrial scale perfusion bioreactor. In addition, CHO-gmt4 cells can be used to produce glycoproteins with mannose-terminated N-glycans. Recombinant glucocerebrosidase produced by CHO-gmt4 cells will not require glycan remodeling and may be directly used to treat patients with Gaucher disease. CHO-gmt4 cells can also be used to produce other glycoprotein therapeutics which target cells expressing mannose receptors.
authors.affiliation: Bioprocessing Technology Institute; Agency for Science, Technology, and Research (A*STAR); Singapore, Singapore.
authors.forename: John S Y
authors.initials: JS
authors.lastname: Goh
authors.affiliation: State Key Laboratory of Bioreactor Engineering; East China University of Science and Technology; Shanghai, China.
authors.forename: Yingwei
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Bioprocessing Technology Institute; Agency for Science, Technology, and Research (A*STAR); Singapore, Singapore.
authors.forename: Kah Fai
authors.initials: KF
authors.lastname: Chan
authors.affiliation: Bioprocessing Technology Institute; Agency for Science, Technology, and Research (A*STAR); Singapore, Singapore.
authors.forename: Corrine
authors.initials: C
authors.lastname: Wan
authors.affiliation: Bioprocessing Technology Institute; Agency for Science, Technology, and Research (A*STAR); Singapore, Singapore.
authors.forename: Gavin
authors.initials: G
authors.lastname: Teo
authors.affiliation: Bioprocessing Technology Institute; Agency for Science, Technology, and Research (A*STAR); Singapore, Singapore.
authors.forename: Peiqing
authors.initials: P
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Bioreactor Engineering; East China University of Science and Technology; Shanghai, China.
authors.forename: Yuanxing
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Bioprocessing Technology Institute; Agency for Science, Technology, and Research (A*STAR); Singapore, Singapore.
authors.forename: Zhiwei
authors.initials: Z
authors.lastname: Song
mesh_terms: D000818:Animals; D060846:Cell Engineering; D004921:Erythropoietin / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D017351:N-Acetylglucosaminyltransferases / Q000378:metabolism*; D019158:N-Acetylneuraminic Acid / Q000378:metabolism*; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics*
chemical_list: D004921:Erythropoietin; D013762:Tetrahydrofolate Dehydrogenase; D017351:N-Acetylglucosaminyltransferases; D019158:N-Acetylneuraminic Acid
keywords: CHO glycosylation mutants; CHO-gmt4; N-acetylglucosaminyltransferase I (GnT I); erythropoietin; mannose-terminated N-glycans; recombinant glycoproteins; sialylation