Combining somatic mutations present in different in vivo affinity-matured antibodies isolated from immunized Lama glama yields ultra-potent antibody therapeutics
| Autor: | Nico Ongenae, Anna Hultberg, Christian Cambillau, Christophe Blanchetot, Torsten Dreier, Silvia Spinelli, Wieger Hemrika, John Wijdenes, Ava Sadi, Rob C. Roovers, Alex Klarenbeek, Anke Kretz-Rommel, Hans de Haard, Georg Schragel, Aline Desmyter |
|---|---|
| Přispěvatelé: | Architecture et fonction des macromolécules biologiques (AFMB), Institut National de la Recherche Agronomique (INRA)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU)-Institut National de la Recherche Agronomique (INRA) |
| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Models Molecular Phage display medicine.drug_class Antibody Affinity Bioengineering Monoclonal antibody Biochemistry 03 medical and health sciences Immunoglobulin Fab Fragments 0302 clinical medicine Antibody Repertoire Peptide Library medicine biology.domesticated_animal Potency Animals Humans Amino Acid Sequence Peptide library Molecular Biology ComputingMilieux_MISCELLANEOUS [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry Molecular Biology/Structural Biology [q-bio.BM] biology Interleukin-6 Models Immunological Molecular biology [SDV.BIBS]Life Sciences [q-bio]/Quantitative Methods [q-bio.QM] Lama glama Recombinant Proteins 3. Good health [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry Molecular Biology/Biomolecules [q-bio.BM] 030104 developmental biology 030220 oncology & carcinogenesis Mutation biology.protein Antibody Camelids New World Sequence Alignment Biotechnology |
| Zdroj: | Protein Engineering, Design and Selection Protein Engineering, Design and Selection, Oxford University Press (OUP), 2016, 29 (4), pp.123-133. ⟨10.1093/protein/gzw003⟩ Protein Engineering, Design and Selection, 2016, 29 (4), pp.123-133. ⟨10.1093/protein/gzw003⟩ |
| ISSN: | 1741-0126 1741-0134 |
| DOI: | 10.1093/protein/gzw003⟩ |
| Popis: | Highly potent human antibodies are required to therapeutically neutralize cytokines such as interleukin-6 (IL-6) that is involved in many inflammatory diseases and malignancies. Although a number of mutagenesis approaches exist to perform antibody affinity maturation, these may cause antibody instability and production issues. Thus, a robust and easy antibody affinity maturation strategy to increase antibody potency remains highly desirable. By immunizing llama, cloning the 'immune' antibody repertoire and using phage display, we selected a diverse set of IL-6 antagonistic Fabs. Heavy chain shuffling was performed on the Fab with lowest off-rate, resulting in a panel of variants with even lower off-rate. Structural analysis of the Fab:IL-6 complex suggests that the increased affinity was partly due to a serine to tyrosine switch in HCDR2. This translated into neutralizing capacity in an in vivo model of IL-6 induced SAA production. Finally, a novel Fab library was designed, encoding all variations found in the natural repertoire of VH genes identified after heavy chain shuffling. High stringency selections resulted in identification of a Fab with 250-fold increased potency when re-formatted into IgG1. Compared with a heavily engineered anti-IL-6 monoclonal antibody currently in clinical development, this IgG was at least equally potent, showing the engineering process to have had led to a highly potent anti-IL-6 antibody. |
| Databáze: | OpenAIRE |
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