Chang, Geen-DongGeen-DongChangChen, Chen-JuChen-JuChenLin, Chih-YinChih-YinLinChen, Hsei-ChornHsei-ChornChenChen, HungwenHungwenChen2009-07-292018-07-062009-07-292018-07-06200301681656http://ntur.lib.ntu.edu.tw//handle/246246/163043The 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.application/pdf359633 bytesapplication/pdfen-USGlycosylation; Glycosyltransferase; Human α1-antitrypsin; Insect cellBlood; 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 baculovirusjournal article10.1016/S0168-1656(02)00364-4126683152-s2.0-12244295475https://www.scopus.com/inward/record.uri?eid=2-s2.0-12244295475&doi=10.1016%2fS0168-1656%2802%2900364-4&partnerID=40&md5=8482c97b1aa818f7fedf1327f06c326fhttp://ntur.lib.ntu.edu.tw/bitstream/246246/163043/1/14.pdf