Journal article
Electrophoretic analysis of glycoprotein glycans produced by lepidopteran insect cells infected with an immediate early recombinant baculovirus encoding mammalian β1,4-galactosyltransferase
Glycoconjugate journal, Vol.16(12), pp.753-756
12/1999
DOI: 10.1023/A:1007131611378
Abstract
Glycosylation, the most extensive co- and post-translational modification of eukaryotic cells, can significantly affect biological activity and is particularly important for recombinant glycoproteins in human therapeutic applications. The baculovirus-insect cell expression system is a popular tool for the expression of heterologous proteins and has an excellent record of producing high levels of biologically active eukaryotic proteins. Insect cells are capable of glycosylation, but their N-glycosylation pathway is truncated in comparison with the pathway of mammalian cells. A previous study demonstrated that an immediate early recombinant baculovirus could be used to extend the insect cell N-glycosylation pathway by contributing bovine β-1,4 galactosyltransferase (GalT) immediately after infection. Lectin blotting assays indicated that this ectopically expressed enzyme could transfer galactose to an N-linked glycan on a foreign glycoprotein expressed later in infection. In the current study, glycans were isolated from total Sf-9 cell glycoproteins after infection with the immediate early recombinant baculovirus encoding GalT, fluorescently conjugated and analyzed by electrophoresis in combination with exoglycosidase digestion. These direct analyses clearly demonstrated that Sf-9 cells infected with this recombinant baculovirus can synthesize galactosylated N-linked glycans.
Details
- Title: Subtitle
- Electrophoretic analysis of glycoprotein glycans produced by lepidopteran insect cells infected with an immediate early recombinant baculovirus encoding mammalian β1,4-galactosyltransferase
- Creators
- Michael Wolff - Division of Medicinal and Natural Products Chemistry, Department of Chemical and Biochemical Engineering University of Wyoming Laramie WY 82071 USADavid Murhammer - Division of Medicinal and Natural Products Chemistry, Department of Chemical and Biochemical Engineering University of Wyoming Laramie WY 82071 USADonald Jarvis - Department of Molecular Biology University of Wyoming Laramie WY 82071 USARobert Linhardt - Department of Chemistry University of Iowa PHAR-S328 Iowa City IA 52242 USA
- Resource Type
- Journal article
- Publication Details
- Glycoconjugate journal, Vol.16(12), pp.753-756
- Publisher
- Kluwer Academic Publishers-Plenum Publishers; New York
- DOI
- 10.1023/A:1007131611378
- ISSN
- 0282-0080
- eISSN
- 1573-4986
- Language
- English
- Date published
- 12/1999
- Academic Unit
- Chemical and Biochemical Engineering
- Record Identifier
- 9984003413402771
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