The functionality of a therapeutic antibody candidate restored by a single mutation from proline to threonine in the variable region
mAbs play an essential role in the therapeutic arsenal. Our laboratory has patented the Rendomab-B49 mAb targeting the endothelin B receptor (ETB). This G protein-coupled receptor plays a driving role in the progression of numerous cancers. We chimerized our mAb (xiRB49) to evaluate its preclinical...
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| Format: | Article |
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Taylor & Francis Group
2023-12-01
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| Series: | Human Vaccines & Immunotherapeutics |
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| Online Access: | https://www.tandfonline.com/doi/10.1080/21645515.2023.2279867 |
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| author | Marie Hautiere Irene Maffucci Narciso Costa Amaury Herbet Sosthene Essono Séverine Padiolleau-Lefevre Didier Boquet |
| author_facet | Marie Hautiere Irene Maffucci Narciso Costa Amaury Herbet Sosthene Essono Séverine Padiolleau-Lefevre Didier Boquet |
| author_sort | Marie Hautiere |
| collection | DOAJ |
| description | mAbs play an essential role in the therapeutic arsenal. Our laboratory has patented the Rendomab-B49 mAb targeting the endothelin B receptor (ETB). This G protein-coupled receptor plays a driving role in the progression of numerous cancers. We chimerized our mAb (xiRB49) to evaluate its preclinical therapeutic efficacy in different ETB+ tumor models with an antibody drug conjugate approach. As previously reported, the chimerization process of an antibody can alter its functionality. In this article, we present the chimerization of RB49. xiRB49 purified by Protein A remained perfectly soluble and did not aggregate, but it lost all its ability to recognize ETB. A detailed analysis of its variable region using IMGT tools allowed us to identify an unusual proline at position 125. In silico mAb modeling and in vitro experiments were performed for a better understanding of xiRB49 structure-function relationships. Our results show that the proline in position 125 on the heavy chain alters the xiRB49 CDR3 light chain conformation and its mutation to threonine allows complete functional recovery. |
| format | Article |
| id | doaj-art-5457d5c48a09474cae6ab11171ce34f8 |
| institution | Kabale University |
| issn | 2164-5515 2164-554X |
| language | English |
| publishDate | 2023-12-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Human Vaccines & Immunotherapeutics |
| spelling | doaj-art-5457d5c48a09474cae6ab11171ce34f82025-08-20T03:26:39ZengTaylor & Francis GroupHuman Vaccines & Immunotherapeutics2164-55152164-554X2023-12-0119310.1080/21645515.2023.2279867The functionality of a therapeutic antibody candidate restored by a single mutation from proline to threonine in the variable regionMarie Hautiere0Irene Maffucci1Narciso Costa2Amaury Herbet3Sosthene Essono4Séverine Padiolleau-Lefevre5Didier Boquet6Département Médicaments et Technologies pour la Santé (DMTS), SPI, Université Paris-Saclay, CEA, Gif-sur-Yvette, FranceCentre de Recherche de Royallieu, CNRS UMR 7025, Génie Enzymatique et Cellulaire, Compiègne Cedex, FranceDépartement Médicaments et Technologies pour la Santé (DMTS), SPI, Université Paris-Saclay, CEA, Gif-sur-Yvette, FranceDépartement Médicaments et Technologies pour la Santé (DMTS), SPI, Université Paris-Saclay, CEA, Gif-sur-Yvette, FranceMedical Biotechnology Engineering LLC, Malden, MA, USACentre de Recherche de Royallieu, CNRS UMR 7025, Génie Enzymatique et Cellulaire, Compiègne Cedex, FranceDépartement Médicaments et Technologies pour la Santé (DMTS), SPI, Université Paris-Saclay, CEA, Gif-sur-Yvette, FrancemAbs play an essential role in the therapeutic arsenal. Our laboratory has patented the Rendomab-B49 mAb targeting the endothelin B receptor (ETB). This G protein-coupled receptor plays a driving role in the progression of numerous cancers. We chimerized our mAb (xiRB49) to evaluate its preclinical therapeutic efficacy in different ETB+ tumor models with an antibody drug conjugate approach. As previously reported, the chimerization process of an antibody can alter its functionality. In this article, we present the chimerization of RB49. xiRB49 purified by Protein A remained perfectly soluble and did not aggregate, but it lost all its ability to recognize ETB. A detailed analysis of its variable region using IMGT tools allowed us to identify an unusual proline at position 125. In silico mAb modeling and in vitro experiments were performed for a better understanding of xiRB49 structure-function relationships. Our results show that the proline in position 125 on the heavy chain alters the xiRB49 CDR3 light chain conformation and its mutation to threonine allows complete functional recovery.https://www.tandfonline.com/doi/10.1080/21645515.2023.2279867Monoclonal antibodychimerizationprolinein silico modelling |
| spellingShingle | Marie Hautiere Irene Maffucci Narciso Costa Amaury Herbet Sosthene Essono Séverine Padiolleau-Lefevre Didier Boquet The functionality of a therapeutic antibody candidate restored by a single mutation from proline to threonine in the variable region Human Vaccines & Immunotherapeutics Monoclonal antibody chimerization proline in silico modelling |
| title | The functionality of a therapeutic antibody candidate restored by a single mutation from proline to threonine in the variable region |
| title_full | The functionality of a therapeutic antibody candidate restored by a single mutation from proline to threonine in the variable region |
| title_fullStr | The functionality of a therapeutic antibody candidate restored by a single mutation from proline to threonine in the variable region |
| title_full_unstemmed | The functionality of a therapeutic antibody candidate restored by a single mutation from proline to threonine in the variable region |
| title_short | The functionality of a therapeutic antibody candidate restored by a single mutation from proline to threonine in the variable region |
| title_sort | functionality of a therapeutic antibody candidate restored by a single mutation from proline to threonine in the variable region |
| topic | Monoclonal antibody chimerization proline in silico modelling |
| url | https://www.tandfonline.com/doi/10.1080/21645515.2023.2279867 |
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