Effect of humanizing mutations on the stability of the llama single-domain variable region
| العنوان: | Effect of humanizing mutations on the stability of the llama single-domain variable region |
|---|---|
| المؤلفون: | Miguel A. Soler, Sara Fortuna, Jiewen Wang, Gregor Bajc, Sandra Folarin Oloketuyi, Ario de Marco, Barbara Medagli, He Huang |
| المساهمون: | Soler, Miguel A., Medagli, Barbara, Wang, Jiewen, Oloketuyi, Sandra, Bajc, Gregor, Huang, He, Fortuna, Sara, Marco, Ario de |
| المصدر: | Biomolecules, vol. 11, no. 2, 163, 2021. Biomolecules Volume 11 Issue 2 Biomolecules, Vol 11, Iss 163, p 163 (2021) |
| بيانات النشر: | MDPI, 2022. |
| سنة النشر: | 2022 |
| مصطلحات موضوعية: | Models, Molecular, In silico, Mutant, lcsh:QR1-502, Heterologous, Enzyme-Linked Immunosorbent Assay, Sequence (biology), Computational biology, Molecular Dynamics Simulation, Biochemistry, lcsh:Microbiology, Article, 03 medical and health sciences, nanobody humanization, Molecular level, Nanobody humanization, Nanobody framework, udc:577, Animals, Cluster Analysis, Humans, Cloning, Molecular, Molecular Biology, CDR grafting, Modeling, Gene Library, 030304 developmental biology, 0303 health sciences, Chemistry, nanobody framework, modeling, 030302 biochemistry & molecular biology, Single-Domain Antibodies, Surface Plasmon Resonance, Antigen binding, Transformation (genetics), Solubility, Mutation, Primary sequence, Camelids, New World, Protein Binding |
| الوصف: | In vivo clinical applications of nanobodies (VHHs) require molecules that induce minimal immunoresponse and therefore possess sequences as similar as possible to the human VH domain. Although the relative sequence variability in llama nanobodies has been used to identify scaffolds with partially humanized signature, the transformation of the Camelidae hallmarks in the framework2 still represents a major problem. We assessed a set of mutants in silico and experimentally to elucidate what is the contribution of single residues to the VHH stability and how their combinations affect the mutant nanobody stability. We described at molecular level how the interaction among residues belonging to different structural elements enabled a model llama nanobody (C8WT, isolated from a naï ve library) to be functional and maintain its stability, despite the analysis of its primary sequence would classify it as aggregation-prone. Five chimeras formed by grafting CDRs isolated from different nanobodies into C8WT scaffold were successfully expressed as soluble proteins and both tested clones preserved their antigen binding specificity. We identified a nanobody with human hallmarks that seems suitable for humanizing selected camelid VHHs by grafting heterologous CDRs in its scaffold and could serve for the preparation of a synthetic library of human-like single domains. |
| وصف الملف: | application/pdf; text/url; ELETTRONICO |
| اللغة: | English |
| تدمد: | 2218-273X |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::13f1cb72bd5bf6e29f070b7e6887c8c4Test https://repozitorij.uni-lj.si/Dokument.php?id=153621&dnTest= |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....13f1cb72bd5bf6e29f070b7e6887c8c4 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 2218273X |
|---|