Antibody humanization—the Influence of the antibody framework on the CDR-H3 loop ensemble in solution
| العنوان: | Antibody humanization—the Influence of the antibody framework on the CDR-H3 loop ensemble in solution |
|---|---|
| المؤلفون: | Monica L. Fernández-Quintero, Klaus R. Liedl, Martin C. Heiss |
| المصدر: | Protein Engineering, Design and Selection |
| بيانات النشر: | Oxford University Press (OUP), 2019. |
| سنة النشر: | 2019 |
| مصطلحات موضوعية: | 0301 basic medicine, conformational selection, humanization, Protein Conformation, medicine.drug_class, Bioengineering, Complementarity determining region, Molecular Dynamics Simulation, immunogenicity, Antibodies, Monoclonal, Humanized, Monoclonal antibody, 01 natural sciences, Biochemistry, 03 medical and health sciences, Molecular dynamics, CDR-H3 loop dynamics, 0103 physical sciences, medicine, Humans, Molecular Biology, 010304 chemical physics, biology, Chemistry, Antigen binding, Complementarity Determining Regions, interdomain dynamics, Solutions, Loop (topology), 030104 developmental biology, biology.protein, Biophysics, Thermodynamics, Original Article, Antibody, Biotechnology |
| الوصف: | Antibody engineering of non-human antibodies has focused on reducing immunogenicity by humanization, being a major limitation in developing monoclonal antibodies. We analyzed four series of antibody binding fragments (Fabs) and a variable fragment (Fv) with structural information in different stages of humanization to investigate the influence of the framework, point mutations and specificity on the complementarity determining region (CDR)-H3 loop dynamics. We also studied a Fv without structural information of the anti-idiotypic antibody Ab2/3H6, because it completely lost its binding affinity upon superhumanization, as an example of a failed humanization. Enhanced sampling techniques in combination with molecular dynamics simulations allow to access micro- to milli-second timescales of the CDR-H3 loop dynamics and reveal kinetic and thermodynamic changes involved in the process of humanization. In most cases, we observe a reduced conformational diversity of the CDR-H3 loop when grafted on a human framework and find a conformational shift of the dominant CDR-H3 loop conformation in solution. A shallow side minimum of the conformational CDR-H3 loop ensemble attached to the murine framework becomes the dominant conformation in solution influenced by the human framework. Additionally, we observe in the case of the failed humanization that the potentially binding competent murine CDR-H3 loop ensemble in solution shows nearly no kinetical or structural overlap with the superhumanized variant, thus explaining the loss of binding. |
| تدمد: | 1741-0134 1741-0126 |
| DOI: | 10.1093/protein/gzaa004 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::8e0c808111de946b84d639e1f88f9826 https://doi.org/10.1093/protein/gzaa004 |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....8e0c808111de946b84d639e1f88f9826 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 17410134 17410126 |
|---|---|
| DOI: | 10.1093/protein/gzaa004 |