Structural basis for improved efficacy of therapeutic antibodies on defucosylation of their Fc glycans

Mizushima, Tsunehiro; Yagi, Hirokazu; Takemoto, Emi; Shibata-Koyama, Mami; Isoda, Yuya; Iida, Shigeru; Masuda, Kouji; Satoh, Mitsuo; Kato, Koichi

doi:10.1111/j.1365-2443.2011.01552.x

Cited by 226 publications

(260 citation statements)

References 34 publications

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“…[12][13][14] The Fc glycan structure of an IgG impacts its effector functions. For example, core-fucosylation has been shown to decrease Fc binding to FcgRIIIa, 6,15,16 which significantly reduces ADCC. 17,18 In addition, the presence of terminal galactose has been shown to induce conformational changes in the Fc domain, 10 increasing Fc binding to C1q which promotes CDC.…”

Section: Introductionmentioning

confidence: 99%

Production of α2,6-sialylated IgG1 in CHO cells

Raymond

Robotham

Spearman

et al. 2015

mAbs

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The presence of a2,6-sialic acids on the Fc N-glycan provides anti-inflammatory properties to the IgGs through a mechanism that remains unclear. Fc-sialylated IgGs are rare in humans as well as in industrial host cell lines such as Chinese hamster ovary (CHO) cells. Facilitated access to well-characterized a2,6-sialylated IgGs would help elucidate the mechanism of this intriguing IgG's effector function. This study presents a method for the efficient Fc glycan a2,6-sialylation of a wild-type and a F243A IgG1 mutant by transient co-expression with the human a2,6-sialyltransferase 1 (ST6) and b1,4-galactosyltransferase 1 (GT) in CHO cells. Overexpression of ST6 alone only had a moderate effect on the glycoprofiles, whereas GT alone greatly enhanced Fc-galactosylation, but not sialylation. Overexpression of both GT and ST6 was necessary to obtain a glycoprofile dominated by a2,6-sialylated glycans in both antibodies. The wild-type was composed of the G2FS(6)1 glycan (38%) with remaining unsialylated glycans, while the mutant glycoprofile was essentially composed of G2FS(6)1 (25%), G2FS(3,6)2 (16%) and G2FS(6,6)2 (37%). The a2,6-linked sialic acids represented over 85% of all sialic acids in both antibodies. We discuss how the limited sialylation level in the wild-type IgG1 expressed alone or with GT results from the glycan interaction with Fc's amino acid residues or from intrinsic galactosyl-and sialyl-transferases substrate specificities.

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Section: Introductionmentioning

confidence: 99%

Production of α2,6-sialylated IgG1 in CHO cells

Raymond

Robotham

Spearman

et al. 2015

mAbs

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“…1 The potential for immunogenicity, i.e., the ability of a biological agent to induce a humoral or cell-mediated immune response, has long been the "Achilles' heel" of therapeutic antibodies. 2,3 When administered, non-human antibodies were commonly recognized as foreign by the immune systems of treated patients, leading to neutralization and rapid clearance of the injected therapeutic antibodies or derivatives thereof, thereby limiting their efficacy. The introduction of chimeric antibodies, in which murine constant domains were replaced by human ones, 4 was an early attempt to reduce immunogenicity.…”

Section: Introductionmentioning

confidence: 99%

Camelid Ig V genes reveal significant human homology not seen in therapeutic target genes, providing for a powerful therapeutic antibody platform

Klarenbeek

Mazouari²,

Desmyter

et al. 2015

mAbs

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de Haard & Ikbel Achour (2015) Camelid Ig V genes reveal significant human homology not seen in therapeutic target genes, providing for a powerful therapeutic antibody platform, mAbs, 7:4, 693-706, DOI: 10.1080/19420862.2015 To link to this article: https://doi.org/10. 1080/19420862.2015 Camelid immunoglobulin variable (IGV) regions were found homologous to their human counterparts; however, the germline V repertoires of camelid heavy and light chains are still incomplete and their therapeutic potential is only beginning to be appreciated. We therefore leveraged the publicly available HTG and WGS databases of Lama pacos and Camelus ferus to retrieve the germline repertoire of V genes using human IGV genes as reference. In addition, we amplified IGKV and IGLV genes to uncover the V germline repertoire of Lama glama and sequenced BAC clones covering part of the Lama pacos IGK and IGL loci. Our in silico analysis showed that camelid counterparts of all human IGKV and IGLV families and most IGHV families could be identified, based on canonical structure and sequence homology. Interestingly, this sequence homology seemed largely restricted to the Ig V genes and was far less apparent in other genes: 6 therapeutically relevant target genes differed significantly from their human orthologs. This contributed to efficient immunization of llamas with the human proteins CD70, MET, interleukin (IL)-1b and IL-6, resulting in large panels of functional antibodies. The in silico predicted human-homologous canonical folds of camelidderived antibodies were confirmed by X-ray crystallography solving the structure of 2 selected camelid anti-CD70 and anti-MET antibodies. These antibodies showed identical fold combinations as found in the corresponding human germline V families, yielding binding site structures closely similar to those occurring in human antibodies. In conclusion, our results indicate that active immunization of camelids can be a powerful therapeutic antibody platform.

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“…In the US, over 20 antibodies using this technique are in clinical phase 1 or 2 trials. The underlying mechanism by which ADCC is highly activated is probably due to the implication of carbohydrate interactions between FcγIIIa receptor and IgG1 (98,99).…”

Section: Core Fucose and Fut8mentioning

confidence: 99%