Combining random mutagenesis, structure-guided design and next-generation sequencing to mitigate polyreactivity of an anti-IL-21R antibody
| Autor: | Virginie McManus, James R. Apgar, Sonia Beyer, Sharon M. Campbell, Orla Cunningham, Matthew Allister Lambert, Joseph DeBartolo, Eric M. Bennett, Lydia Mosyak |
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| Rok vydání: | 2021 |
| Předmět: |
Protein Conformation
Immunology Mutagenesis (molecular biology technique) Computational biology Protein Engineering Antibody nonspecificity DNA sequencing 03 medical and health sciences Structure-Activity Relationship 0302 clinical medicine Drug Stability In vivo Antibody Specificity Report developability Immunology and Allergy Humans 030304 developmental biology 0303 health sciences biology Protein Stability High-Throughput Nucleotide Sequencing Interleukin-21 Receptor alpha Subunit Antibodies Neutralizing HEK293 Cells Mutagenesis 030220 oncology & carcinogenesis Drug Design biology.protein Computer-Aided Design Antibody deselection Platform development optimization Reports IL-21R |
| Zdroj: | mAbs article-version (VoR) Version of Record |
| ISSN: | 1942-0870 |
| Popis: | Despite substantial technological advances in antibody library and display platform development, the number of approved biotherapeutics from displayed libraries remains limited. In vivo, 20–50% of peripheral B cells undergo a process of receptor editing, which modifies the variable and junctional regions of light chains to delete auto-reactive clones. However, in vitro antibody evolution relies primarily on interaction with antigen, with no in-built checkpoints to ensure that the selected antibodies have not acquired additional specificities or biophysical liabilities during the optimization process. We had previously observed an enrichment of positive charge in the complementarity-determining regions of an anti-IL-21 R antibody during affinity optimization, which correlated with more potent IL-21 neutralization, but poor in vivo pharmacokinetics (PK). There is an emerging body of data that has correlated antibody nonspecificity with poor PK in vivo, and established a series of screening assays that are predictive of this behavior. In this study we revisit the challenge of developing an anti-IL-21 R antibody that can effectively compete with IL-21 for its highly negatively charged paratope while maintaining favorable biophysical properties. In vitro deselection methods that included an excess of negatively charged membrane preparations, or deoxyribonucleic acid, during phage selection of optimization libraries were unsuccessful in avoiding enrichment of highly charged, nonspecific antibody variants. However, a combination of structure-guided rational library design, next-generation sequencing of library outputs and application of linear regression models resulted in the identification of an antibody that maintained high affinity for IL-21 R and exhibited a desirable stability and biophysical profile. |
| Databáze: | OpenAIRE |
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