1. Influence of canonical structure determining residues on antibody affinity and stability.
- Author
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Clark LA, Demarest SJ, Eldredge J, Jarpe MB, Li Y, Simon K, and van Vlijmen HW
- Subjects
- Amino Acid Substitution, Animals, Antibody Affinity, Binding Sites, Complementarity Determining Regions genetics, Humans, Hydrogen Bonding, Immunoglobulin Heavy Chains genetics, Immunoglobulin Light Chains genetics, Integrin alpha1beta1 chemistry, Integrin alpha1beta1 immunology, Models, Molecular, Protein Binding, Protein Engineering, Protein Interaction Domains and Motifs, Protein Stability, Protein Unfolding, Rats, Thermodynamics, Complementarity Determining Regions chemistry, Immunoglobulin Heavy Chains chemistry, Immunoglobulin Light Chains chemistry
- Abstract
A number of light and heavy chain canonical residue core redesigns were made in a therapeutic antibody (AQC2, anti-VLA1) Fab to explore the consequences to binding affinity and stability. These positions are all loop supporting, primarily CDR1 residues which do not directly contact the antigen. Structure based methods were used with and without consensus sequence information. 30 constructs were made, 24 expressed, and 70% of the designs using consensus sequence information retained binding affinity. Some success maintaining stability with more extreme redesigns suggests a surprising tolerance to mutation, though it often comes at the cost of loss of binding affinity and presumed loop conformation changes. In concordance with the expected need to present an ordered surface for binding, a relationship between decreased affinity and decreased stability was observed. Overpacking the core tends to destabilize the molecule and should be avoided., (Copyright © 2013 Elsevier Inc. All rights reserved.)
- Published
- 2014
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