Sialylation of human IgG-Fc carbohydrate by transfected rat alpha2,6- sialyltransferase

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Sialylation of human IgG-Fc carbohydrate by transfected rat alpha2,6- sialyltransferase. / Jassal, R; Jenkins, N; Charlwood, J; Camilleri, P; Jefferis, Royston; Lund, John.

In: Biochemical and Biophysical Research Communications, Vol. 286, No. 2, 17.08.2001, p. 243-9.

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@article{b8714dde1a044886b61f110c6c43c8e2,
title = "Sialylation of human IgG-Fc carbohydrate by transfected rat alpha2,6- sialyltransferase",
abstract = "A recombinant IgG3 antibody with Phe-243 replaced by Ala (FA243) was expressed in a CHO-K1 parental cell line. The resulting IgG-Fc-linked carbohydrate was significantly alpha2,3-sialylated (53% of glycans), as indicated by normal- and reverse-phase HPLC analyses. Following transfection of a rat alpha2,6-sialyltransferase gene into this parental cell line, IgG-Fc-linked glycans were sialylated (60% of glycans) such that the ratio of alpha2,6- to alpha2,3-linked sialic acid was 0.9:1.0. By comparison, the wild-type IgG3 (F243) is minimally sialylated (2-3% alpha2,3-linked), thus suggesting that sialylation is controlled primarily by the protein structure local to the carbohydrate and that the two sialyltransferases compete to sialylate the nascent oligosaccharide. The additional alpha2,6-sialylation affected the function of the recombinant antibody. FA243 IgG3 having both alpha2,6 and alpha2,3-sialylation restored recognition to wild-type IgG3 levels for human FcgammaRI, FcgammaRII, and target cell lysis by complement. We discuss how sialylation linkage could modulate IgG function.",
keywords = "glycosylation, complement, sialylation, Fc gamma receptors, IgG",
author = "R Jassal and N Jenkins and J Charlwood and P Camilleri and Royston Jefferis and John Lund",
year = "2001",
month = aug,
day = "17",
doi = "10.1006/bbrc.2001.5382",
language = "English",
volume = "286",
pages = "243--9",
journal = "Biochemical and Biophysical Research Communications",
issn = "0006-291X",
publisher = "Elsevier",
number = "2",

}

RIS

TY - JOUR

T1 - Sialylation of human IgG-Fc carbohydrate by transfected rat alpha2,6- sialyltransferase

AU - Jassal, R

AU - Jenkins, N

AU - Charlwood, J

AU - Camilleri, P

AU - Jefferis, Royston

AU - Lund, John

PY - 2001/8/17

Y1 - 2001/8/17

N2 - A recombinant IgG3 antibody with Phe-243 replaced by Ala (FA243) was expressed in a CHO-K1 parental cell line. The resulting IgG-Fc-linked carbohydrate was significantly alpha2,3-sialylated (53% of glycans), as indicated by normal- and reverse-phase HPLC analyses. Following transfection of a rat alpha2,6-sialyltransferase gene into this parental cell line, IgG-Fc-linked glycans were sialylated (60% of glycans) such that the ratio of alpha2,6- to alpha2,3-linked sialic acid was 0.9:1.0. By comparison, the wild-type IgG3 (F243) is minimally sialylated (2-3% alpha2,3-linked), thus suggesting that sialylation is controlled primarily by the protein structure local to the carbohydrate and that the two sialyltransferases compete to sialylate the nascent oligosaccharide. The additional alpha2,6-sialylation affected the function of the recombinant antibody. FA243 IgG3 having both alpha2,6 and alpha2,3-sialylation restored recognition to wild-type IgG3 levels for human FcgammaRI, FcgammaRII, and target cell lysis by complement. We discuss how sialylation linkage could modulate IgG function.

AB - A recombinant IgG3 antibody with Phe-243 replaced by Ala (FA243) was expressed in a CHO-K1 parental cell line. The resulting IgG-Fc-linked carbohydrate was significantly alpha2,3-sialylated (53% of glycans), as indicated by normal- and reverse-phase HPLC analyses. Following transfection of a rat alpha2,6-sialyltransferase gene into this parental cell line, IgG-Fc-linked glycans were sialylated (60% of glycans) such that the ratio of alpha2,6- to alpha2,3-linked sialic acid was 0.9:1.0. By comparison, the wild-type IgG3 (F243) is minimally sialylated (2-3% alpha2,3-linked), thus suggesting that sialylation is controlled primarily by the protein structure local to the carbohydrate and that the two sialyltransferases compete to sialylate the nascent oligosaccharide. The additional alpha2,6-sialylation affected the function of the recombinant antibody. FA243 IgG3 having both alpha2,6 and alpha2,3-sialylation restored recognition to wild-type IgG3 levels for human FcgammaRI, FcgammaRII, and target cell lysis by complement. We discuss how sialylation linkage could modulate IgG function.

KW - glycosylation

KW - complement

KW - sialylation

KW - Fc gamma receptors

KW - IgG

UR - http://www.scopus.com/inward/record.url?scp=0034815962&partnerID=8YFLogxK

U2 - 10.1006/bbrc.2001.5382

DO - 10.1006/bbrc.2001.5382

M3 - Article

C2 - 11500028

VL - 286

SP - 243

EP - 249

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 0006-291X

IS - 2

ER -