Affinity enhancement of an in vivo matured therapeutic antibody using structure-based computational design
| Autor: | Stefano F. Liparoto, John Eldredge, You Li, Christopher Fitch, Mia Rushe, Alexey Lugovskoy, Karl J. M. Hanf, Matthew Jarpe, Herman W. T. van Vlijmen, Woody Sherman, Kenneth Simon, Louis A. Clark, P. Ann Boriack-Sjodin, Miller Stephan S, Bethany Friedman |
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| Rok vydání: | 2006 |
| Předmět: |
Models
Molecular Antigen-Antibody Complex Mutant Antibody Affinity Immunoglobulins Mutagenesis (molecular biology technique) Crystallography X-Ray Biochemistry Article Antibodies Integrin alpha1beta1 Protein–protein interaction Affinity maturation Structure-Activity Relationship Structure–activity relationship Molecular Biology chemistry.chemical_classification Hydrogen bond Biomolecule Crystallography Amino Acid Substitution chemistry Drug Design Biophysics Computer-Aided Design Binding Sites Antibody |
| Zdroj: | Protein Science. 15:949-960 |
| ISSN: | 1469-896X 0961-8368 |
| Popis: | Improving the affinity of a high-affinity protein-protein interaction is a challenging problem that has practical applications in the development of therapeutic biomolecules. We used a combination of structure-based computational methods to optimize the binding affinity of an antibody fragment to the I-domain of the integrin VLA1. Despite the already high affinity of the antibody (Kd approximately 7 nM) and the moderate resolution (2.8 A) of the starting crystal structure, the affinity was increased by an order of magnitude primarily through a decrease in the dissociation rate. We determined the crystal structure of a high-affinity quadruple mutant complex at 2.2 A. The structure shows that the design makes the predicted contacts. Structural evidence and mutagenesis experiments that probe a hydrogen bond network illustrate the importance of satisfying hydrogen bonding requirements while seeking higher-affinity mutations. The large and diverse set of interface mutations allowed refinement of the mutant binding affinity prediction protocol and improvement of the single-mutant success rate. Our results indicate that structure-based computational design can be successfully applied to further improve the binding of high-affinity antibodies. |
| Databáze: | OpenAIRE |
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