https://scholars.lib.ntu.edu.tw/handle/123456789/143565
標題: | Improvement of glycosylation in insect cells with mammalian glycosyltransferases | 作者: | Chang, Geen-Dong Chen, Chen-Ju Lin, Chih-Yin Chen, Hsei-Chorn Chen, Hungwen |
關鍵字: | Glycosylation; Glycosyltransferase; Human α1-antitrypsin; Insect cell | 公開日期: | 2003 | 起(迄)頁: | 61-71 | 來源出版物: | Journal of Biotechnology | 摘要: | The N-glycans of recombinant glycoproteins expressed in insect cells mainly contain high mannose or tri-mannose structures, which are truncated forms of the sialylated N-glycans found in mammalian cells. Because asialylated glycoproteins have a shorter half-life in blood circulation, we investigated if sialylated therapeutic glycoprotein can be produced from insect cells by enhancing the N-glycosylation machinery of the cells. We co-expressed in two insect cell lines, Sf9 and Ea4, the human α1-antitrypsin (hα1AT) protein with a series of key glycosyltransferases, including GlcNAc transferase II (GnT2), β1,4-galactosyltransferase (β14GT), and α2,6-sialyltransferase (α26ST) by a single recombinant baculovirus. We demonstrated that the enhancement of N-glycosylation is cell type-dependent and is more efficient in Ea4 than Sf9 cells. Glycan analysis indicated that sialylated hα1AT proteins were produced in Ea4 insect cells expressing the above-mentioned exogenous glycosyltransferases. Therefore, our expression strategy may simplify the production of humanized therapeutic glycoproteins by improving the N-glycosylation pathway in specific insect cells, with an ensemble of exogenous glycosyltransferases in a single recombinant baculovirus. © 2003 Elsevier Science B.V. All rights reserved. |
URI: | http://ntur.lib.ntu.edu.tw//handle/246246/163043 | ISSN: | 01681656 | DOI: | 10.1016/S0168-1656(02)00364-4 | SDG/關鍵字: | Blood; Cells; Proteins; Viruses; Insect cells; Genetic engineering; alpha 1 antitrypsin; alpha 2,6 sialyltransferase; beta 1,4 galactosyltransferase; glycosyltransferase; n acetylglucosaminyltransferase; sialoglycoprotein; unclassified drug; article; Baculovirus; cell line; cell type; controlled study; enzyme glycosylation; expression vector; insect cell; mammal; nonhuman; priority journal; protein expression; virus recombinant; Insecta; Mammalia; unidentified baculovirus |
顯示於: | 生化科學研究所 |
在 IR 系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。