Profiling the Biophysical Developability Properties of Common IgG1 Fc Effector Silencing Variants
Therapeutic antibodies represent the most significant modality in biologics, with around 150 approved drugs on the market. In addition to specific target binding mediated by the variable fragments (Fvs) of the heavy and light chains, antibodies possess effector functions through binding of the const...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-08-01
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| Series: | Antibodies |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2073-4468/12/3/54 |
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| author | Robert Pejchal Anthony B. Cooper Michael E. Brown Maximiliano Vásquez Eric M. Krauland |
| author_facet | Robert Pejchal Anthony B. Cooper Michael E. Brown Maximiliano Vásquez Eric M. Krauland |
| author_sort | Robert Pejchal |
| collection | DOAJ |
| description | Therapeutic antibodies represent the most significant modality in biologics, with around 150 approved drugs on the market. In addition to specific target binding mediated by the variable fragments (Fvs) of the heavy and light chains, antibodies possess effector functions through binding of the constant region (Fc) to Fcγ receptors (FcγR), which allow immune cells to attack and kill target cells using a variety of mechanisms. However, for some applications, including T-cell-engaging bispecifics, this effector function is typically undesired. Mutations within the lower hinge and the second constant domain (CH2) of IgG1 that comprise the FcγR binding interface reduce or eliminate effector function (“Fc silencing”) while retaining binding to the neonatal Fc receptor (FcRn), important for normal antibody pharmacokinetics (PKs). Comprehensive profiling of biophysical developability properties would benefit the choice of constant region variants for development. Here, we produce a large panel of representative mutations previously described in the literature and in many cases in clinical or approved molecules, generate select combinations thereof, and characterize their binding and biophysical properties. We find that some commonly used CH2 mutations, including D265A and P331S, are effective in reducing binding to FcγR but significantly reduce stability, promoting aggregation, particularly under acidic conditions commonly employed in manufacturing. We highlight mutation sets that are particularly effective for eliminating Fc effector function with the retention of WT-like stability, including L234A, L235A, and S267K (LALA-S267K), L234A, L235E, and S267K (LALE-S267K), L234A, L235A, and P329A (LALA-P329A), and L234A, L235E, and P329G (LALE-P329G). |
| first_indexed | 2024-03-10T23:06:36Z |
| format | Article |
| id | doaj.art-88e7dc5c418f4819ba70208472531bf6 |
| institution | Directory Open Access Journal |
| issn | 2073-4468 |
| language | English |
| last_indexed | 2024-03-10T23:06:36Z |
| publishDate | 2023-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Antibodies |
| spelling | doaj.art-88e7dc5c418f4819ba70208472531bf62023-11-19T09:18:26ZengMDPI AGAntibodies2073-44682023-08-011235410.3390/antib12030054Profiling the Biophysical Developability Properties of Common IgG1 Fc Effector Silencing VariantsRobert Pejchal0Anthony B. Cooper1Michael E. Brown2Maximiliano Vásquez3Eric M. Krauland4Adimab LLC, Lebanon, NH 03766, USAAclys Bio, Lebanon, NH 03766, USAAdimab LLC, Lebanon, NH 03766, USAAdimab LLC, Lebanon, NH 03766, USAAdimab LLC, Lebanon, NH 03766, USATherapeutic antibodies represent the most significant modality in biologics, with around 150 approved drugs on the market. In addition to specific target binding mediated by the variable fragments (Fvs) of the heavy and light chains, antibodies possess effector functions through binding of the constant region (Fc) to Fcγ receptors (FcγR), which allow immune cells to attack and kill target cells using a variety of mechanisms. However, for some applications, including T-cell-engaging bispecifics, this effector function is typically undesired. Mutations within the lower hinge and the second constant domain (CH2) of IgG1 that comprise the FcγR binding interface reduce or eliminate effector function (“Fc silencing”) while retaining binding to the neonatal Fc receptor (FcRn), important for normal antibody pharmacokinetics (PKs). Comprehensive profiling of biophysical developability properties would benefit the choice of constant region variants for development. Here, we produce a large panel of representative mutations previously described in the literature and in many cases in clinical or approved molecules, generate select combinations thereof, and characterize their binding and biophysical properties. We find that some commonly used CH2 mutations, including D265A and P331S, are effective in reducing binding to FcγR but significantly reduce stability, promoting aggregation, particularly under acidic conditions commonly employed in manufacturing. We highlight mutation sets that are particularly effective for eliminating Fc effector function with the retention of WT-like stability, including L234A, L235A, and S267K (LALA-S267K), L234A, L235E, and S267K (LALE-S267K), L234A, L235A, and P329A (LALA-P329A), and L234A, L235E, and P329G (LALE-P329G).https://www.mdpi.com/2073-4468/12/3/54antibodyCD3T-cell engagerFcγ receptorFc effector silencingC1q |
| spellingShingle | Robert Pejchal Anthony B. Cooper Michael E. Brown Maximiliano Vásquez Eric M. Krauland Profiling the Biophysical Developability Properties of Common IgG1 Fc Effector Silencing Variants Antibodies antibody CD3 T-cell engager Fcγ receptor Fc effector silencing C1q |
| title | Profiling the Biophysical Developability Properties of Common IgG1 Fc Effector Silencing Variants |
| title_full | Profiling the Biophysical Developability Properties of Common IgG1 Fc Effector Silencing Variants |
| title_fullStr | Profiling the Biophysical Developability Properties of Common IgG1 Fc Effector Silencing Variants |
| title_full_unstemmed | Profiling the Biophysical Developability Properties of Common IgG1 Fc Effector Silencing Variants |
| title_short | Profiling the Biophysical Developability Properties of Common IgG1 Fc Effector Silencing Variants |
| title_sort | profiling the biophysical developability properties of common igg1 fc effector silencing variants |
| topic | antibody CD3 T-cell engager Fcγ receptor Fc effector silencing C1q |
| url | https://www.mdpi.com/2073-4468/12/3/54 |
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