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Glycobiology Advance Access originally published online on November 1, 2002
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Glycobiology, 2003, Vol. 13, No. 1 23-34
© 2003 Oxford University Press

Complex-type biantennary N-glycans of recombinant human transferrin from Trichoplusia ni insect cells expressing mammalian ß-1,4-galactosyltransferase and ß-1,2-N-acetylglucosaminyltransferase II

Noboru Tomiya12, Dale Howe3, Jared J. Aumiller3, Manuj Pathak4, Jung Park5, Karen B. Palter5, Donald L. Jarvis3, Michael J. Betenbaugh4 and Yuan C. Lee2

2 Department of Biology, Johns Hopkins University, 3400 North Charles Street, Baltimore, MD 21218, USA
3 Department of Molecular Biology, University of Wyoming, P.o. Box 3944, Laramie, WY 82071-3944, USA
4 Department of Chemical Engineering, Johns Hopkins University,3400 North Charles Street, Baltimore, MD 21218, USA
5 Department of Biology, Temple University, 1900 North 12th Street, Philadelphia, PA 19122, USA

Received on July 21, 2002; revised on August 29, 2002; accepted on September 5, 2002

A novel recombinant baculovirus expression vector was used to produce His-tagged human transferrin in a transformed insect cell line (Tn5ß4GalT) that constitutively expresses a mammalian ß-1,4-galactosyltransferase. This virus encoded the His-tagged human transferrin protein in conventional fashion under the control of the very late polyhedrin promoter. In addition, to enhance the synthesis of galactosylated biantennary N-glycans, this virus encoded human ß-1,2- N-acetylglucosaminyltransferase II under the control of an immediate-early (ie1) promoter. Detailed analyses by MALDI-TOF MS, exoglycosidase digestion, and two-dimensional HPLC revealed that the N-glycans on the purified recombinant human transferrin produced by this virus–host system included four different fully galactosylated, biantennary, complex-type glycans. Thus, this study describes a novel baculovirus–host system, which can be used to produce a recombinant glycoprotein with fully galactosylated, biantennary N-glycans.

1 To whom correspondence should be addressed;e-mail: ntomiya1{at}jhu.edu


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O. Choi, N. Tomiya, J. H. Kim, J. M. Slavicek, M. J. Betenbaugh, and Y. C. Lee
N-glycan structures of human transferrin produced by Lymantria dispar (gypsy moth) cells using the LdMNPV expression system
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