Discovery of a single-subunit oligosaccharyltransferase that enables glycosylation of full-length IgG antibodies in bacteria
Abstract Human immunoglobulin G (IgG) antibodies are a major class of biotherapeutics and undergo N-linked glycosylation in their Fc domain, which is critical for immune functions and therapeutic activity. Hence, technologies for producing authentically glycosylated IgGs are in high demand. Previous...
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Nature Portfolio
2025-07-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-025-61440-7 |
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| author | Belen Sotomayor Thomas C. Donahue Sai Pooja Mahajan May N. Taw Sophia W. Hulbert Erik J. Bidstrup D. Natasha Owitipana Alexandra Pang Xu Yang Souvik Ghosal Christopher A. Alabi Parastoo Azadi Jeffrey J. Gray Michael C. Jewett Lai-Xi Wang Matthew P. DeLisa |
| author_facet | Belen Sotomayor Thomas C. Donahue Sai Pooja Mahajan May N. Taw Sophia W. Hulbert Erik J. Bidstrup D. Natasha Owitipana Alexandra Pang Xu Yang Souvik Ghosal Christopher A. Alabi Parastoo Azadi Jeffrey J. Gray Michael C. Jewett Lai-Xi Wang Matthew P. DeLisa |
| author_sort | Belen Sotomayor |
| collection | DOAJ |
| description | Abstract Human immunoglobulin G (IgG) antibodies are a major class of biotherapeutics and undergo N-linked glycosylation in their Fc domain, which is critical for immune functions and therapeutic activity. Hence, technologies for producing authentically glycosylated IgGs are in high demand. Previous attempts to engineer Escherichia coli for this purpose have met limited success due in part to the lack of oligosaccharyltransferase (OST) enzymes that can install N-glycans at the conserved N297 site in the Fc region. Here, we identify a single-subunit OST from Desulfovibrio marinus with relaxed substrate specificity that catalyzes glycosylation of native Fc acceptor sites. By chemoenzymatic remodeling the attached bacterial glycans to homogeneous, asialo complex-type G2 N-glycans, the E. coli-derived Fc binds human FcγRIIIa/CD16a, a key receptor for antibody-dependent cellular cytotoxicity (ADCC). Overall, the discovery of D. marinus OST provides previously unavailable biocatalytic capabilities and sets the stage for using E. coli to produce fully human antibodies. |
| format | Article |
| id | doaj-art-d16444c84b5d4da4ab58e453ee04e884 |
| institution | DOAJ |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-d16444c84b5d4da4ab58e453ee04e8842025-08-20T03:03:37ZengNature PortfolioNature Communications2041-17232025-07-0116111610.1038/s41467-025-61440-7Discovery of a single-subunit oligosaccharyltransferase that enables glycosylation of full-length IgG antibodies in bacteriaBelen Sotomayor0Thomas C. Donahue1Sai Pooja Mahajan2May N. Taw3Sophia W. Hulbert4Erik J. Bidstrup5D. Natasha Owitipana6Alexandra Pang7Xu Yang8Souvik Ghosal9Christopher A. Alabi10Parastoo Azadi11Jeffrey J. Gray12Michael C. Jewett13Lai-Xi Wang14Matthew P. DeLisa15Robert Frederick Smith School of Chemical and Biomolecular Engineering, Cornell UniversityRobert Frederick Smith School of Chemical and Biomolecular Engineering, Cornell UniversityDepartment of Chemical and Biomolecular Engineering, Johns Hopkins UniversityDepartment of Microbiology, Cornell UniversityBiochemistry, Molecular and Cell Biology, Cornell UniversityRobert Frederick Smith School of Chemical and Biomolecular Engineering, Cornell UniversityDepartment of Chemistry and Biochemistry, University of MarylandRobert Frederick Smith School of Chemical and Biomolecular Engineering, Cornell UniversityComplex Carbohydrate Research Center, University of Georgia, 315 Riverbend RoadDepartment of Chemistry and Chemical Biology, Cornell UniversityRobert Frederick Smith School of Chemical and Biomolecular Engineering, Cornell UniversityComplex Carbohydrate Research Center, University of Georgia, 315 Riverbend RoadDepartment of Chemical and Biomolecular Engineering, Johns Hopkins UniversityDepartment of Bioengineering, Stanford UniversityDepartment of Chemistry and Biochemistry, University of MarylandRobert Frederick Smith School of Chemical and Biomolecular Engineering, Cornell UniversityAbstract Human immunoglobulin G (IgG) antibodies are a major class of biotherapeutics and undergo N-linked glycosylation in their Fc domain, which is critical for immune functions and therapeutic activity. Hence, technologies for producing authentically glycosylated IgGs are in high demand. Previous attempts to engineer Escherichia coli for this purpose have met limited success due in part to the lack of oligosaccharyltransferase (OST) enzymes that can install N-glycans at the conserved N297 site in the Fc region. Here, we identify a single-subunit OST from Desulfovibrio marinus with relaxed substrate specificity that catalyzes glycosylation of native Fc acceptor sites. By chemoenzymatic remodeling the attached bacterial glycans to homogeneous, asialo complex-type G2 N-glycans, the E. coli-derived Fc binds human FcγRIIIa/CD16a, a key receptor for antibody-dependent cellular cytotoxicity (ADCC). Overall, the discovery of D. marinus OST provides previously unavailable biocatalytic capabilities and sets the stage for using E. coli to produce fully human antibodies.https://doi.org/10.1038/s41467-025-61440-7 |
| spellingShingle | Belen Sotomayor Thomas C. Donahue Sai Pooja Mahajan May N. Taw Sophia W. Hulbert Erik J. Bidstrup D. Natasha Owitipana Alexandra Pang Xu Yang Souvik Ghosal Christopher A. Alabi Parastoo Azadi Jeffrey J. Gray Michael C. Jewett Lai-Xi Wang Matthew P. DeLisa Discovery of a single-subunit oligosaccharyltransferase that enables glycosylation of full-length IgG antibodies in bacteria Nature Communications |
| title | Discovery of a single-subunit oligosaccharyltransferase that enables glycosylation of full-length IgG antibodies in bacteria |
| title_full | Discovery of a single-subunit oligosaccharyltransferase that enables glycosylation of full-length IgG antibodies in bacteria |
| title_fullStr | Discovery of a single-subunit oligosaccharyltransferase that enables glycosylation of full-length IgG antibodies in bacteria |
| title_full_unstemmed | Discovery of a single-subunit oligosaccharyltransferase that enables glycosylation of full-length IgG antibodies in bacteria |
| title_short | Discovery of a single-subunit oligosaccharyltransferase that enables glycosylation of full-length IgG antibodies in bacteria |
| title_sort | discovery of a single subunit oligosaccharyltransferase that enables glycosylation of full length igg antibodies in bacteria |
| url | https://doi.org/10.1038/s41467-025-61440-7 |
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