Structural analysis of light chain-driven bispecific antibodies targeting CD47 and PD-L1
In contrast to natural antibodies that rely mainly on the heavy chain to establish contacts with their cognate antigen, we have developed a bispecific antibody format in which the light chain (LC) drives antigen binding and specificity. To better understand epitope-paratope interactions in this cont...
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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/10.1080/19420862.2024.2362432 |
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| author | Pauline Malinge Xavier Chauchet Jérémie Bourguignon Nicolas Bosson Sébastien Calloud Tereza Bautzova Marie Borlet Mette Laursen Vinardas Kelpsas Nadia Rose Franck Gueneau Ulla Ravn Giovanni Magistrelli Nicolas Fischer |
| author_facet | Pauline Malinge Xavier Chauchet Jérémie Bourguignon Nicolas Bosson Sébastien Calloud Tereza Bautzova Marie Borlet Mette Laursen Vinardas Kelpsas Nadia Rose Franck Gueneau Ulla Ravn Giovanni Magistrelli Nicolas Fischer |
| author_sort | Pauline Malinge |
| collection | DOAJ |
| description | In contrast to natural antibodies that rely mainly on the heavy chain to establish contacts with their cognate antigen, we have developed a bispecific antibody format in which the light chain (LC) drives antigen binding and specificity. To better understand epitope-paratope interactions in this context, we determined the X-ray crystallographic structures of an antigen binding fragment (Fab) in complex with human CD47 and another Fab in complex with human PD-L1. These Fabs contain a κ-LC and a λ-LC, respectively, which are paired with an identical heavy chain (HC). The structural analysis of these complexes revealed the dominant contribution of the LCs to antigen binding, but also that the common HC provides some contacts in both CD47 and PD-L1 Fab complexes. The anti-CD47 Fab was affinity optimized by diversifying complementary-determining regions of the LC followed by phage display selections. Using homology modeling, the contributions of the amino acid modification to the affinity increase were analyzed. Our results demonstrate that, despite a less prominent role in natural antibodies, the LC can mediate high affinity binding to different antigens and neutralize their biological function. Importantly, Fabs containing a common variable heavy (VH) domain enable the generation of bispecific antibodies retaining a truly native structure, maximizing their therapeutic potential. |
| format | Article |
| id | doaj-art-5417c41908304d66b55eaa0280c419ce |
| 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-5417c41908304d66b55eaa0280c419ce2025-01-31T04:19:38ZengTaylor & Francis GroupmAbs1942-08621942-08702024-12-0116110.1080/19420862.2024.2362432Structural analysis of light chain-driven bispecific antibodies targeting CD47 and PD-L1Pauline Malinge0Xavier Chauchet1Jérémie Bourguignon2Nicolas Bosson3Sébastien Calloud4Tereza Bautzova5Marie Borlet6Mette Laursen7Vinardas Kelpsas8Nadia Rose9Franck Gueneau10Ulla Ravn11Giovanni Magistrelli12Nicolas Fischer13Light Chain Bioscience - Novimmune SA, Plan-les-Ouates, SwitzerlandLight Chain Bioscience - Novimmune SA, Plan-les-Ouates, SwitzerlandLight Chain Bioscience - Novimmune SA, Plan-les-Ouates, SwitzerlandLight Chain Bioscience - Novimmune SA, Plan-les-Ouates, SwitzerlandLight Chain Bioscience - Novimmune SA, Plan-les-Ouates, SwitzerlandLight Chain Bioscience - Novimmune SA, Plan-les-Ouates, SwitzerlandLight Chain Bioscience - Novimmune SA, Plan-les-Ouates, SwitzerlandSARomics Biostructures AB, Lund, SwedenSARomics Biostructures AB, Lund, SwedenSARomics Biostructures AB, Lund, SwedenLight Chain Bioscience - Novimmune SA, Plan-les-Ouates, SwitzerlandLight Chain Bioscience - Novimmune SA, Plan-les-Ouates, SwitzerlandLight Chain Bioscience - Novimmune SA, Plan-les-Ouates, SwitzerlandLight Chain Bioscience - Novimmune SA, Plan-les-Ouates, SwitzerlandIn contrast to natural antibodies that rely mainly on the heavy chain to establish contacts with their cognate antigen, we have developed a bispecific antibody format in which the light chain (LC) drives antigen binding and specificity. To better understand epitope-paratope interactions in this context, we determined the X-ray crystallographic structures of an antigen binding fragment (Fab) in complex with human CD47 and another Fab in complex with human PD-L1. These Fabs contain a κ-LC and a λ-LC, respectively, which are paired with an identical heavy chain (HC). The structural analysis of these complexes revealed the dominant contribution of the LCs to antigen binding, but also that the common HC provides some contacts in both CD47 and PD-L1 Fab complexes. The anti-CD47 Fab was affinity optimized by diversifying complementary-determining regions of the LC followed by phage display selections. Using homology modeling, the contributions of the amino acid modification to the affinity increase were analyzed. Our results demonstrate that, despite a less prominent role in natural antibodies, the LC can mediate high affinity binding to different antigens and neutralize their biological function. Importantly, Fabs containing a common variable heavy (VH) domain enable the generation of bispecific antibodies retaining a truly native structure, maximizing their therapeutic potential.https://www.tandfonline.com/doi/10.1080/19420862.2024.2362432Bispecificlight chainCD47PD-L1structureX-ray crystallography |
| spellingShingle | Pauline Malinge Xavier Chauchet Jérémie Bourguignon Nicolas Bosson Sébastien Calloud Tereza Bautzova Marie Borlet Mette Laursen Vinardas Kelpsas Nadia Rose Franck Gueneau Ulla Ravn Giovanni Magistrelli Nicolas Fischer Structural analysis of light chain-driven bispecific antibodies targeting CD47 and PD-L1 mAbs Bispecific light chain CD47 PD-L1 structure X-ray crystallography |
| title | Structural analysis of light chain-driven bispecific antibodies targeting CD47 and PD-L1 |
| title_full | Structural analysis of light chain-driven bispecific antibodies targeting CD47 and PD-L1 |
| title_fullStr | Structural analysis of light chain-driven bispecific antibodies targeting CD47 and PD-L1 |
| title_full_unstemmed | Structural analysis of light chain-driven bispecific antibodies targeting CD47 and PD-L1 |
| title_short | Structural analysis of light chain-driven bispecific antibodies targeting CD47 and PD-L1 |
| title_sort | structural analysis of light chain driven bispecific antibodies targeting cd47 and pd l1 |
| topic | Bispecific light chain CD47 PD-L1 structure X-ray crystallography |
| url | https://www.tandfonline.com/doi/10.1080/19420862.2024.2362432 |
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