An efficient system for bioconjugation based on a widely applicable engineered O-glycosylation tag Short title: Controlled bioconjugation via O-glycan engineering
| Autor: | Thomas V. Murray, Kasia Kozakowska-McDonnell, Adam Tibbles, Annabel Taylor, Daniel Higazi, Emmanuel Rossy, Alessandra Rossi, Sivaneswary Genapathy, Giulia Tamburrino, Nicola Rath, Natalie Tigue, Vivian Lindo, Tristan Vaughan, Monika A. Papworth |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: | |
| Zdroj: | mAbs, Vol 13, Iss 1 (2021) |
| Druh dokumentu: | article |
| ISSN: | 19420862 1942-0870 1942-0862 |
| DOI: | 10.1080/19420862.2021.1992068 |
| Popis: | Bioconjugates are an important class of therapeutic molecules. To date, O-glycan-based metabolic glycoengineering has had limited use in this field, due to the complexities of the endogenous O-glycosylation pathway and the lack of an O-glycosylation consensus sequence. Here, we describe the development of a versatile on-demand O-glycosylation system that uses a novel, widely applicable 5 amino acid O-glycosylation tag, and a metabolically engineered UDP-galactose-4-eperimase (GALE) knock-out cell line. Optimization of the primary sequence of the tag enables the production of Fc-based proteins with either single or multiple O-glycans with complexity fully controlled by media supplementation. We demonstrate how the uniformly labeled proteins containing exclusively N-azido-acetylgalactosamine are used for CLICK chemistry-based bioconjugation to generate site-specifically fluorochrome-labeled antibodies, dual-payload molecules, and bioactive Fc-peptides for applications in basic research and drug discovery. To our knowledge, this is the first description of generating a site-specific O-glycosylation system by combining an O-glycosylation tag and a metabolically engineered cell line. |
| Databáze: | Directory of Open Access Journals |
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