| RRC ID |
38913
|
| 著者 |
Mimura Y, Kelly RM, Unwin L, Albrecht S, Jefferis R, Goodall M, Mizukami Y, Mimura-Kimura Y, Matsumoto T, Ueoka H, Rudd PM.
|
| タイトル |
Enhanced sialylation of a human chimeric IgG1 variant produced in human and rodent cell lines.
|
| ジャーナル |
J Immunol Methods
|
| 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.
|
| 巻・号 |
428
|
| ページ |
30-6
|
| 公開日 |
2016-1-1
|
| DOI |
10.1016/j.jim.2015.11.009
|
| PII |
S0022-1759(15)30064-8
|
| PMID |
26627984
|
| MeSH |
Animals
CHO Cells
Cell Line, Tumor
Chromatography, High Pressure Liquid
Cricetulus
Glycosylation
HEK293 Cells
Humans
Immunoglobulin G / biosynthesis
Immunoglobulin G / chemistry
Immunoglobulin G / genetics*
Immunoglobulin G / metabolism*
Mice
Mice, Inbred BALB C
N-Acetylneuraminic Acid / analysis
N-Acetylneuraminic Acid / metabolism*
|
| IF |
1.901
|
| 引用数 |
23
|
|
WOS 分野
|
BIOCHEMICAL RESEARCH METHODS
IMMUNOLOGY
|
| リソース情報 |
| ヒト・動物細胞 |
293(RCB1637) |
