Variable domain mutational analysis to probe the molecular mechanisms of high viscosity of an IgG1 antibody
Subcutaneous injection is the preferred route of administration for many antibody therapeutics for reasons that include its speed and convenience. However, the small volume limit (typically [Formula: see text]2 mL) for subcutaneous delivery often necessitates antibody formulations at high concentrat...
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| Format: | Article |
| Language: | English |
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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/10.1080/19420862.2024.2304282 |
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| author | Jing Dai Saeed Izadi Jonathan Zarzar Patrick Wu Angela Oh Paul J. Carter |
| author_facet | Jing Dai Saeed Izadi Jonathan Zarzar Patrick Wu Angela Oh Paul J. Carter |
| author_sort | Jing Dai |
| collection | DOAJ |
| description | Subcutaneous injection is the preferred route of administration for many antibody therapeutics for reasons that include its speed and convenience. However, the small volume limit (typically [Formula: see text]2 mL) for subcutaneous delivery often necessitates antibody formulations at high concentrations (commonly ≥100 mg/mL), which may lead to physicochemical problems. For example, antibodies with large hydrophobic or charged patches can be prone to self-interaction giving rise to high viscosity. Here, we combined X-ray crystallography with computational modeling to predict regions of an anti-glucagon receptor (GCGR) IgG1 antibody prone to self-interaction. An extensive mutational analysis was undertaken of the complementarity-determining region residues residing in hydrophobic surface patches predicted by spatial aggregation propensity, in conjunction with residue-level solvent accessibility, averaged over conformational ensembles from molecular dynamics simulations. Dynamic light scattering (DLS) was used as a medium throughput screen for self-interaction of ~ 200 anti-GCGR IgG1 variants. A negative correlation was found between the viscosity determined at high concentration (180 mg/mL) and the DLS interaction parameter measured at low concentration (2–10 mg/mL). Additionally, anti-GCGR variants were readily identified with reduced viscosity and antigen-binding affinity within a few fold of the parent antibody, with no identified impact on overall developability. The methods described here may be useful in the optimization of other antibodies to facilitate their therapeutic administration at high concentration. |
| format | Article |
| id | doaj-art-a50ad440254f4b97bb89e51aeaf46a98 |
| 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-a50ad440254f4b97bb89e51aeaf46a982025-01-31T04:19:38ZengTaylor & Francis GroupmAbs1942-08621942-08702024-12-0116110.1080/19420862.2024.2304282Variable domain mutational analysis to probe the molecular mechanisms of high viscosity of an IgG1 antibodyJing Dai0Saeed Izadi1Jonathan Zarzar2Patrick Wu3Angela Oh4Paul J. Carter5Department 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 Bioanalytical Sciences, Genentech, Inc, South San Francisco, CA, USADepartment of Structural Biology, Genentech, Inc, South San Francisco, CA, USADepartment of Antibody Engineering, Genentech, Inc, South San Francisco, CA, USASubcutaneous injection is the preferred route of administration for many antibody therapeutics for reasons that include its speed and convenience. However, the small volume limit (typically [Formula: see text]2 mL) for subcutaneous delivery often necessitates antibody formulations at high concentrations (commonly ≥100 mg/mL), which may lead to physicochemical problems. For example, antibodies with large hydrophobic or charged patches can be prone to self-interaction giving rise to high viscosity. Here, we combined X-ray crystallography with computational modeling to predict regions of an anti-glucagon receptor (GCGR) IgG1 antibody prone to self-interaction. An extensive mutational analysis was undertaken of the complementarity-determining region residues residing in hydrophobic surface patches predicted by spatial aggregation propensity, in conjunction with residue-level solvent accessibility, averaged over conformational ensembles from molecular dynamics simulations. Dynamic light scattering (DLS) was used as a medium throughput screen for self-interaction of ~ 200 anti-GCGR IgG1 variants. A negative correlation was found between the viscosity determined at high concentration (180 mg/mL) and the DLS interaction parameter measured at low concentration (2–10 mg/mL). Additionally, anti-GCGR variants were readily identified with reduced viscosity and antigen-binding affinity within a few fold of the parent antibody, with no identified impact on overall developability. The methods described here may be useful in the optimization of other antibodies to facilitate their therapeutic administration at high concentration.https://www.tandfonline.com/doi/10.1080/19420862.2024.2304282Antibodyintermolecular interactionsmulti-parameter optimizationreversible self-associationrheologysubcutaneous delivery |
| spellingShingle | Jing Dai Saeed Izadi Jonathan Zarzar Patrick Wu Angela Oh Paul J. Carter Variable domain mutational analysis to probe the molecular mechanisms of high viscosity of an IgG1 antibody mAbs Antibody intermolecular interactions multi-parameter optimization reversible self-association rheology subcutaneous delivery |
| title | Variable domain mutational analysis to probe the molecular mechanisms of high viscosity of an IgG1 antibody |
| title_full | Variable domain mutational analysis to probe the molecular mechanisms of high viscosity of an IgG1 antibody |
| title_fullStr | Variable domain mutational analysis to probe the molecular mechanisms of high viscosity of an IgG1 antibody |
| title_full_unstemmed | Variable domain mutational analysis to probe the molecular mechanisms of high viscosity of an IgG1 antibody |
| title_short | Variable domain mutational analysis to probe the molecular mechanisms of high viscosity of an IgG1 antibody |
| title_sort | variable domain mutational analysis to probe the molecular mechanisms of high viscosity of an igg1 antibody |
| topic | Antibody intermolecular interactions multi-parameter optimization reversible self-association rheology subcutaneous delivery |
| url | https://www.tandfonline.com/doi/10.1080/19420862.2024.2304282 |
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