Site-specific glycosylation of an aglycosylated human IgG1-Fc antibody protein generates neoglycocoprotein with enhanced function

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@article{732e28d5cbfb4fe9aba9534370a4fddd,
title = "Site-specific glycosylation of an aglycosylated human IgG1-Fc antibody protein generates neoglycocoprotein with enhanced function",
abstract = "A range of well-defined IgG glycoforms was prepared by employing a combination of synthetic carbohydrate chemistry and genetic engineering. The key aspect of this methodology is the coupling of thioaldoses with cysteine-containing proteins to give disulfide-linked neoglycoproteins. This technology was applied to the synthesis of a series of synthetic N-glycan thioaldoses which were coupled to an aglycosylated IgG1-Fc fragment, engineered to have Cys-297 in place of glycan-linked Asn (Deltah-Fc N297C). Analysis of the resulting Fc neoglycoproteins by mass spectrometry and trypsin digestion showed that the saccharides were site-selectively incorporated at Cys-297 to full occupancy without affecting other Fc protein disulfides. The neoglycoproteins were tested for their ability to interact with human FcgammaRI by inhibiting superoxide production by gamma-interferon-stimulated U937 cells. The neoglycoproteins displayed enhanced superoxide inhibition relative to aglycosylated Deltah-Fc N297C, where increased glycan size correlated positively with increased inhibition.",
author = "GM Watt and John Lund and Jacqueline Levens and K Kolli and Royston Jefferis and G-J Boons",
year = "2003",
month = sep,
day = "1",
doi = "10.1016/j.chembiol.2003.08.006",
language = "English",
volume = "10",
pages = "807--814",
journal = "Chemistry & Biology",
issn = "1074-5521",
publisher = "Elsevier",
number = "9",

}

RIS

TY - JOUR

T1 - Site-specific glycosylation of an aglycosylated human IgG1-Fc antibody protein generates neoglycocoprotein with enhanced function

AU - Watt, GM

AU - Lund, John

AU - Levens, Jacqueline

AU - Kolli, K

AU - Jefferis, Royston

AU - Boons, G-J

PY - 2003/9/1

Y1 - 2003/9/1

N2 - A range of well-defined IgG glycoforms was prepared by employing a combination of synthetic carbohydrate chemistry and genetic engineering. The key aspect of this methodology is the coupling of thioaldoses with cysteine-containing proteins to give disulfide-linked neoglycoproteins. This technology was applied to the synthesis of a series of synthetic N-glycan thioaldoses which were coupled to an aglycosylated IgG1-Fc fragment, engineered to have Cys-297 in place of glycan-linked Asn (Deltah-Fc N297C). Analysis of the resulting Fc neoglycoproteins by mass spectrometry and trypsin digestion showed that the saccharides were site-selectively incorporated at Cys-297 to full occupancy without affecting other Fc protein disulfides. The neoglycoproteins were tested for their ability to interact with human FcgammaRI by inhibiting superoxide production by gamma-interferon-stimulated U937 cells. The neoglycoproteins displayed enhanced superoxide inhibition relative to aglycosylated Deltah-Fc N297C, where increased glycan size correlated positively with increased inhibition.

AB - A range of well-defined IgG glycoforms was prepared by employing a combination of synthetic carbohydrate chemistry and genetic engineering. The key aspect of this methodology is the coupling of thioaldoses with cysteine-containing proteins to give disulfide-linked neoglycoproteins. This technology was applied to the synthesis of a series of synthetic N-glycan thioaldoses which were coupled to an aglycosylated IgG1-Fc fragment, engineered to have Cys-297 in place of glycan-linked Asn (Deltah-Fc N297C). Analysis of the resulting Fc neoglycoproteins by mass spectrometry and trypsin digestion showed that the saccharides were site-selectively incorporated at Cys-297 to full occupancy without affecting other Fc protein disulfides. The neoglycoproteins were tested for their ability to interact with human FcgammaRI by inhibiting superoxide production by gamma-interferon-stimulated U937 cells. The neoglycoproteins displayed enhanced superoxide inhibition relative to aglycosylated Deltah-Fc N297C, where increased glycan size correlated positively with increased inhibition.

UR - http://www.scopus.com/inward/record.url?scp=0141565183&partnerID=8YFLogxK

U2 - 10.1016/j.chembiol.2003.08.006

DO - 10.1016/j.chembiol.2003.08.006

M3 - Article

C2 - 14522051

VL - 10

SP - 807

EP - 814

JO - Chemistry & Biology

JF - Chemistry & Biology

SN - 1074-5521

IS - 9

ER -