| Téléchargement | - Voir le manuscrit accepté : Production of α2,6-sialylated IgG1 in CHO cells (PDF, 1.1 Mio)
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| DOI | Trouver le DOI : https://doi.org/10.1080/19420862.2015.1029215 |
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| Auteur | Rechercher : Raymond, Céline1; Rechercher : Robotham, Anna1; Rechercher : Spearman, Maureen; Rechercher : Butler, Michael; Rechercher : Kelly, John1; Rechercher : Durocher, Yves1 |
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| Affiliation | - Conseil national de recherches du Canada. Thérapeutique en santé humaine
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| Format | Texte, Article |
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| Sujet | B4GALT1; CHO cells; ECL, Erythrina Cristagalli lectin; GT, β1,4-galactosyltransferase 1; HILIC, hydrophilic interaction liquid chromatography; IgG1; LC-ESI-MS, liquid chromatography coupled to electrospray ionization mass spectrometry; MAL-II, Maackia Amurensis lectin II; N-glycosylation; PEI, polyethylenimine; SIAT1; SNA, Sambucus Nigra agglutinin; ST6, α2,6-sialyltransferase 1; TZM, trastuzumab (Herceptin®); cIEF, capillary zone electrophoresis isoelectric focusing; mAbs, monoclonal antibodies; sialylation; transfection; α2,3SA, α2,3-linked sialic acid; α2,6SA, α2,6-linked sialic acid |
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| Résumé | The presence of α2,6-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 α2,6-sialylated IgGs would help elucidate the mechanism of this intriguing IgG's effector function. This study presents a method for the efficient Fc glycan α2,6-sialylation of a wild-type and a F243A IgG1 mutant by transient co-expression with the human α2,6-sialyltransferase 1 (ST6) and β1,4-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 α2,6-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 α2,6-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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| Date de publication | 2015-04-15 |
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| Maison d’édition | Taylor & Francis Group |
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| Dans | |
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| Langue | anglais |
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| Publications évaluées par des pairs | Oui |
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| Numéro du CNRC | NRC-HHT-53284 |
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| Numéro NPARC | 21275405 |
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| Exporter la notice | Exporter en format RIS |
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| Signaler une correction | Signaler une correction (s'ouvre dans un nouvel onglet) |
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| Identificateur de l’enregistrement | e6369b2f-f442-4fe0-96b1-6e446ebe188f |
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| Enregistrement créé | 2015-07-08 |
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| Enregistrement modifié | 2020-06-04 |
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