Improvement of glycosylation in insect cells with mammalian glycosyltransferases
Resource
Journal of Biotechnology 102 (1): 61-71
Journal
Journal of Biotechnology
Pages
61-71
Date Issued
2003
Date
2003
Author(s)
Abstract
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.
Subjects
Glycosylation; Glycosyltransferase; Human α1-antitrypsin; Insect cell
Other Subjects
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
Type
journal article
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