RRC ID 38913
Author Mimura Y, Kelly RM, Unwin L, Albrecht S, Jefferis R, Goodall M, Mizukami Y, Mimura-Kimura Y, Matsumoto T, Ueoka H, Rudd PM.
Title Enhanced sialylation of a human chimeric IgG1 variant produced in human and rodent cell lines.
Journal 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.
Volume 428
Pages 30-6
Published 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
Times Cited 23
Human and Animal Cells 293(RCB1637)