Modulation of the high concentration viscosity of IgG1 antibodies using clinically validated Fc mutations
The self-association of therapeutic antibodies can result in elevated viscosity and create problems in manufacturing and formulation, as well as limit delivery by subcutaneous injection. The high concentration viscosity of some antibodies has been reduced by variable domain mutations or by the addit...
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Taylor & Francis Group
2024-12-01
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| Series: | mAbs |
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| Online Access: | https://www.tandfonline.com/doi/10.1080/19420862.2024.2379560 |
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| author | Joel Heisler Daniel Kovner Saeed Izadi Jonathan Zarzar Paul J. Carter |
| author_facet | Joel Heisler Daniel Kovner Saeed Izadi Jonathan Zarzar Paul J. Carter |
| author_sort | Joel Heisler |
| collection | DOAJ |
| description | The self-association of therapeutic antibodies can result in elevated viscosity and create problems in manufacturing and formulation, as well as limit delivery by subcutaneous injection. The high concentration viscosity of some antibodies has been reduced by variable domain mutations or by the addition of formulation excipients. In contrast, the impact of Fc mutations on antibody viscosity has been minimally explored. Here, we studied the effect of a panel of common and clinically validated Fc mutations on the viscosity of two closely related humanized IgG1, κ antibodies, omalizumab (anti-IgE) and trastuzumab (anti-HER2). Data presented here suggest that both Fab-Fab and Fab-Fc interactions contribute to the high viscosity of omalizumab, in a four-contact model of self-association. Most strikingly, the high viscosity of omalizumab (176 cP) was reduced 10.7- and 2.2-fold by Fc modifications for half-life extension (M252Y:S254T:T256E) and aglycosylation (N297G), respectively. Related single mutations (S254T and T256E) each reduced the viscosity of omalizumab by ~6-fold. An alternative half-life extension Fc mutant (M428L:N434S) had the opposite effect in increasing the viscosity of omalizumab by 1.5-fold. The low viscosity of trastuzumab (8.6 cP) was unchanged or increased by [Formula: see text]2-fold by the different Fc variants. Molecular dynamics simulations provided mechanistic insight into the impact of Fc mutations in modulating electrostatic and hydrophobic surface properties as well as conformational stability of the Fc. This study demonstrates that high viscosity of some IgG1 antibodies can be mitigated by Fc mutations, and thereby offers an additional tool to help design future antibody therapeutics potentially suitable for subcutaneous delivery. |
| format | Article |
| id | doaj-art-1c2c74ea9eeb426685561bfdd47cda1b |
| institution | Kabale University |
| issn | 1942-0862 1942-0870 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | mAbs |
| spelling | doaj-art-1c2c74ea9eeb426685561bfdd47cda1b2025-01-31T04:19:38ZengTaylor & Francis GroupmAbs1942-08621942-08702024-12-0116110.1080/19420862.2024.2379560Modulation of the high concentration viscosity of IgG1 antibodies using clinically validated Fc mutationsJoel Heisler0Daniel Kovner1Saeed Izadi2Jonathan Zarzar3Paul J. Carter4Department of Antibody Engineering, Genentech, Inc, South San Francisco, CA, USADepartment of Pharmaceutical Development, Genentech, Inc, South San Francisco, CA, USADepartment of Pharmaceutical Development, Genentech, Inc, South San Francisco, CA, USADepartment of Pharmaceutical Development, Genentech, Inc, South San Francisco, CA, USADepartment of Antibody Engineering, Genentech, Inc, South San Francisco, CA, USAThe self-association of therapeutic antibodies can result in elevated viscosity and create problems in manufacturing and formulation, as well as limit delivery by subcutaneous injection. The high concentration viscosity of some antibodies has been reduced by variable domain mutations or by the addition of formulation excipients. In contrast, the impact of Fc mutations on antibody viscosity has been minimally explored. Here, we studied the effect of a panel of common and clinically validated Fc mutations on the viscosity of two closely related humanized IgG1, κ antibodies, omalizumab (anti-IgE) and trastuzumab (anti-HER2). Data presented here suggest that both Fab-Fab and Fab-Fc interactions contribute to the high viscosity of omalizumab, in a four-contact model of self-association. Most strikingly, the high viscosity of omalizumab (176 cP) was reduced 10.7- and 2.2-fold by Fc modifications for half-life extension (M252Y:S254T:T256E) and aglycosylation (N297G), respectively. Related single mutations (S254T and T256E) each reduced the viscosity of omalizumab by ~6-fold. An alternative half-life extension Fc mutant (M428L:N434S) had the opposite effect in increasing the viscosity of omalizumab by 1.5-fold. The low viscosity of trastuzumab (8.6 cP) was unchanged or increased by [Formula: see text]2-fold by the different Fc variants. Molecular dynamics simulations provided mechanistic insight into the impact of Fc mutations in modulating electrostatic and hydrophobic surface properties as well as conformational stability of the Fc. This study demonstrates that high viscosity of some IgG1 antibodies can be mitigated by Fc mutations, and thereby offers an additional tool to help design future antibody therapeutics potentially suitable for subcutaneous delivery.https://www.tandfonline.com/doi/10.1080/19420862.2024.2379560Intermolecular interactionsintramolecular interactionsmolecular dynamicsrheologyself-associationsubcutaneous delivery |
| spellingShingle | Joel Heisler Daniel Kovner Saeed Izadi Jonathan Zarzar Paul J. Carter Modulation of the high concentration viscosity of IgG1 antibodies using clinically validated Fc mutations mAbs Intermolecular interactions intramolecular interactions molecular dynamics rheology self-association subcutaneous delivery |
| title | Modulation of the high concentration viscosity of IgG1 antibodies using clinically validated Fc mutations |
| title_full | Modulation of the high concentration viscosity of IgG1 antibodies using clinically validated Fc mutations |
| title_fullStr | Modulation of the high concentration viscosity of IgG1 antibodies using clinically validated Fc mutations |
| title_full_unstemmed | Modulation of the high concentration viscosity of IgG1 antibodies using clinically validated Fc mutations |
| title_short | Modulation of the high concentration viscosity of IgG1 antibodies using clinically validated Fc mutations |
| title_sort | modulation of the high concentration viscosity of igg1 antibodies using clinically validated fc mutations |
| topic | Intermolecular interactions intramolecular interactions molecular dynamics rheology self-association subcutaneous delivery |
| url | https://www.tandfonline.com/doi/10.1080/19420862.2024.2379560 |
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