Rational affinity maturation of anti-amyloid antibodies with high conformational and sequence specificity
Autor: | Jennifer M. Zupancic, Shannon J. Moore, Henry L. Paulson, Alexandra B. Sutter, Edward Ionescu, Julia E. Gerson, Magdalena I. Ivanova, Yulei Zhang, Geoffrey G. Murphy, Peter M. Tessier, Matthew D. Smith, Alec A. Desai, Charles G. Starr, Emily K. Makowski |
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Rok vydání: | 2020 |
Předmět: |
0301 basic medicine
Models Molecular yeast surface display Protein Conformation Aβ amyloid beta Protein aggregation Biochemistry scFv Mice directed evolution biology antibody engineering Chemistry fibril neurodegeneration Antibodies Monoclonal Brain ELISA enzyme-linked immunosorbent assay 3. Good health aggregate Aducanumab Research Article Amyloid Amyloid beta medicine.drug_class VH variable domain of heavy chain Monoclonal antibody Affinity maturation 03 medical and health sciences CDR complementarity-determining region medicine Single-chain variable fragment Animals Humans Molecular Biology mAb 030102 biochemistry & molecular biology scFv single-chain variable fragment VL variable domain of light chain Cell Biology 030104 developmental biology Case-Control Studies Crenezumab biology.protein Alzheimer BSA bovine serum albumin Binding Sites Antibody Fc fragment crystallizable |
Zdroj: | The Journal of Biological Chemistry |
ISSN: | 1083-351X |
Popis: | The aggregation of amyloidogenic polypeptides is strongly linked to several neurodegenerative disorders, including Alzheimer's and Parkinson's diseases. Conformational antibodies that selectively recognize protein aggregates are leading therapeutic agents for selectively neutralizing toxic aggregates, diagnostic and imaging agents for detecting disease, and biomedical reagents for elucidating disease mechanisms. Despite their importance, it is challenging to generate high-quality conformational antibodies in a systematic and site-specific manner due to the properties of protein aggregates (hydrophobic, multivalent, and heterogeneous) and limitations of immunization (uncontrolled antigen presentation and immunodominant epitopes). Toward addressing these challenges, we have developed a systematic directed evolution procedure for affinity maturing antibodies against Alzheimer's Aβ fibrils and selecting variants with strict conformational and sequence specificity. We first designed a library based on a lead conformational antibody by sampling combinations of amino acids in the antigen-binding site predicted to mediate high antibody specificity. Next, we displayed this library on the surface of yeast, sorted it against Aβ42 aggregates, and identified promising clones using deep sequencing. The resulting antibodies displayed similar or higher affinities than clinical-stage Aβ antibodies (aducanumab and crenezumab). Moreover, the affinity-matured antibodies retained high conformational specificity for Aβ aggregates, as observed for aducanumab and unlike crenezumab. Notably, the affinity-maturated antibodies displayed extremely low levels of nonspecific interactions, as observed for crenezumab and unlike aducanumab. We expect that our systematic methods for generating antibodies with unique combinations of desirable properties will improve the generation of high-quality conformational antibodies specific for diverse types of aggregated conformers. |
Databáze: | OpenAIRE |
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