Enhanced sialylation of a human chimeric IgG1 variant produced in human and rodent cell lines

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Enhanced sialylation of a human chimeric IgG1 variant produced in human and rodent cell lines. / Mimura, Yusuke; Kelly, Ronan M.; Unwin, Louise; Albrecht, Simone; Jefferis, Royston; Goodall, Margaret; Mizukami, Yoichi; Mimura-kimura, Yuka; Matsumoto, Tsuneo; Ueoka, Hiroshi; Rudd, Pauline M.

In: Journal of Immunological Methods, Vol. 428, 01.2016, p. 30-36.

Research output: Contribution to journalArticlepeer-review

Harvard

Mimura, Y, Kelly, RM, Unwin, L, Albrecht, S, Jefferis, R, Goodall, M, Mizukami, Y, Mimura-kimura, Y, Matsumoto, T, Ueoka, H & Rudd, PM 2016, 'Enhanced sialylation of a human chimeric IgG1 variant produced in human and rodent cell lines', Journal of Immunological Methods, vol. 428, pp. 30-36. https://doi.org/10.1016/j.jim.2015.11.009

APA

Mimura, Y., Kelly, R. M., Unwin, L., Albrecht, S., Jefferis, R., Goodall, M., Mizukami, Y., Mimura-kimura, Y., Matsumoto, T., Ueoka, H., & Rudd, P. M. (2016). Enhanced sialylation of a human chimeric IgG1 variant produced in human and rodent cell lines. Journal of Immunological Methods, 428, 30-36. https://doi.org/10.1016/j.jim.2015.11.009

Vancouver

Author

Mimura, Yusuke ; Kelly, Ronan M. ; Unwin, Louise ; Albrecht, Simone ; Jefferis, Royston ; Goodall, Margaret ; Mizukami, Yoichi ; Mimura-kimura, Yuka ; Matsumoto, Tsuneo ; Ueoka, Hiroshi ; Rudd, Pauline M. / Enhanced sialylation of a human chimeric IgG1 variant produced in human and rodent cell lines. In: Journal of Immunological Methods. 2016 ; Vol. 428. pp. 30-36.

Bibtex

@article{5e3300e3acb1480988c4b9daf6426b8e,
title = "Enhanced sialylation of a human chimeric IgG1 variant produced in human and rodent cell lines",
abstract = "Glycosylation of the IgG-Fc is essential for optimal binding and activation of Fcγ receptors and the C1q component of complement. However, it has been reported that the effector functions are down-regulated when the Fc glycans terminate in sialic acid residues and that sialylated IgG mediates anti-inflammatory effects of intravenous immunoglobulin (IVIG). Although recombinant IgG is hypo-sialylated, Fc sialylation is shown to be markedly increased when a mouse/human chimeric IgG3 Phe243Ala (F243A) variant is expressed in Chinese hamster ovary (CHO)-K1 cells. Here we investigate whether sialylation is increased in IgG1 F243A when expressed in CHO-K1, mouse myeloma J558L and human embryonic kidney (HEK) 293. Although the sialylation level was 2–5% for IgG1 wild type (WT), it was increased to 31%, 10% and 33% for the variant from CHO-K1, J558L and HEK293 cells, respectively. Interestingly, an increased addition of bisecting GlcNAc and α(1-3)-galactose residues to the Fc glycan was observed for HEK293-derived and J558L-derived IgG1 F243A, respectively. Fucosylation of HEK293-derived IgG1 F243A was maintained despite increased bisecting GlcNAc content. Although sialic acid and bisecting GlcNAc residues are reported to have an opposing effect on antibody-dependent cellular cytotoxicity (ADCC), IgG1 F243A showed 7 times lower ADCC activities than IgG1 WT, irrespective of bisecting GlcNAc residue. Thus, highly sialylated, human cell-derived IgG1 F243A with lowered ADCC activity may be of interest for the development of therapeutic antibodies with anti-inflammatory properties as an alternative to IVIG.",
keywords = "Glycosylation, IgG, Intravenous immunoglobulin, Sialic acid, Therapeutic antibody",
author = "Yusuke Mimura and Kelly, {Ronan M.} and Louise Unwin and Simone Albrecht and Royston Jefferis and Margaret Goodall and Yoichi Mizukami and Yuka Mimura-kimura and Tsuneo Matsumoto and Hiroshi Ueoka and Rudd, {Pauline M.}",
year = "2016",
month = jan,
doi = "10.1016/j.jim.2015.11.009",
language = "English",
volume = "428",
pages = "30--36",
journal = "Journal of Immunological Methods",
issn = "0022-1759",
publisher = "Elsevier",

}

RIS

TY - JOUR

T1 - Enhanced sialylation of a human chimeric IgG1 variant produced in human and rodent cell lines

AU - Mimura, Yusuke

AU - Kelly, Ronan M.

AU - Unwin, Louise

AU - Albrecht, Simone

AU - Jefferis, Royston

AU - Goodall, Margaret

AU - Mizukami, Yoichi

AU - Mimura-kimura, Yuka

AU - Matsumoto, Tsuneo

AU - Ueoka, Hiroshi

AU - Rudd, Pauline M.

PY - 2016/1

Y1 - 2016/1

N2 - Glycosylation of the IgG-Fc is essential for optimal binding and activation of Fcγ receptors and the C1q component of complement. However, it has been reported that the effector functions are down-regulated when the Fc glycans terminate in sialic acid residues and that sialylated IgG mediates anti-inflammatory effects of intravenous immunoglobulin (IVIG). Although recombinant IgG is hypo-sialylated, Fc sialylation is shown to be markedly increased when a mouse/human chimeric IgG3 Phe243Ala (F243A) variant is expressed in Chinese hamster ovary (CHO)-K1 cells. Here we investigate whether sialylation is increased in IgG1 F243A when expressed in CHO-K1, mouse myeloma J558L and human embryonic kidney (HEK) 293. Although the sialylation level was 2–5% for IgG1 wild type (WT), it was increased to 31%, 10% and 33% for the variant from CHO-K1, J558L and HEK293 cells, respectively. Interestingly, an increased addition of bisecting GlcNAc and α(1-3)-galactose residues to the Fc glycan was observed for HEK293-derived and J558L-derived IgG1 F243A, respectively. Fucosylation of HEK293-derived IgG1 F243A was maintained despite increased bisecting GlcNAc content. Although sialic acid and bisecting GlcNAc residues are reported to have an opposing effect on antibody-dependent cellular cytotoxicity (ADCC), IgG1 F243A showed 7 times lower ADCC activities than IgG1 WT, irrespective of bisecting GlcNAc residue. Thus, highly sialylated, human cell-derived IgG1 F243A with lowered ADCC activity may be of interest for the development of therapeutic antibodies with anti-inflammatory properties as an alternative to IVIG.

AB - Glycosylation of the IgG-Fc is essential for optimal binding and activation of Fcγ receptors and the C1q component of complement. However, it has been reported that the effector functions are down-regulated when the Fc glycans terminate in sialic acid residues and that sialylated IgG mediates anti-inflammatory effects of intravenous immunoglobulin (IVIG). Although recombinant IgG is hypo-sialylated, Fc sialylation is shown to be markedly increased when a mouse/human chimeric IgG3 Phe243Ala (F243A) variant is expressed in Chinese hamster ovary (CHO)-K1 cells. Here we investigate whether sialylation is increased in IgG1 F243A when expressed in CHO-K1, mouse myeloma J558L and human embryonic kidney (HEK) 293. Although the sialylation level was 2–5% for IgG1 wild type (WT), it was increased to 31%, 10% and 33% for the variant from CHO-K1, J558L and HEK293 cells, respectively. Interestingly, an increased addition of bisecting GlcNAc and α(1-3)-galactose residues to the Fc glycan was observed for HEK293-derived and J558L-derived IgG1 F243A, respectively. Fucosylation of HEK293-derived IgG1 F243A was maintained despite increased bisecting GlcNAc content. Although sialic acid and bisecting GlcNAc residues are reported to have an opposing effect on antibody-dependent cellular cytotoxicity (ADCC), IgG1 F243A showed 7 times lower ADCC activities than IgG1 WT, irrespective of bisecting GlcNAc residue. Thus, highly sialylated, human cell-derived IgG1 F243A with lowered ADCC activity may be of interest for the development of therapeutic antibodies with anti-inflammatory properties as an alternative to IVIG.

KW - Glycosylation

KW - IgG

KW - Intravenous immunoglobulin

KW - Sialic acid

KW - Therapeutic antibody

U2 - 10.1016/j.jim.2015.11.009

DO - 10.1016/j.jim.2015.11.009

M3 - Article

VL - 428

SP - 30

EP - 36

JO - Journal of Immunological Methods

JF - Journal of Immunological Methods

SN - 0022-1759

ER -