1. Structure-Based Design of Supercharged, Highly Thermoresistant Antibodies
- Author
-
Supriya Pai, Alena M Calm, Heather Welsh, Candice Warner, Christien Kluwe, James Carney, Aroop Sircar, R. E. Hughes, Vlad Codrea, Jianqing Xu, Aleksandr E. Miklos, Patricia E. Buckley, Brian Kuhlman, Monica Berrondo, Andrew D. Ellington, George Georgiou, Bryan S. Der, Jeffrey J. Gray, and Melody Zacharko
- Subjects
Protein Folding ,Clinical Biochemistry ,Protein Engineering ,Biochemistry ,Article ,Drug Discovery ,Homology modeling ,Molecular Biology ,chemistry.chemical_classification ,Pharmacology ,biology ,Chemistry ,Temperature ,Hydrogen Bonding ,General Medicine ,Protein engineering ,Combinatorial chemistry ,Protein Structure, Tertiary ,Amino acid ,Mutation (genetic algorithm) ,biology.protein ,Biophysics ,Structure based ,Molecular Medicine ,Protein folding ,Antibody ,Software ,Single-Chain Antibodies - Abstract
Summary Mutation of surface residues to charged amino acids increases resistance to aggregation and can enable reversible unfolding. We have developed a protocol using the Rosetta computational design package that "supercharges" proteins while considering the energetic implications of each mutation. Using a homology model, a single-chain variable fragment antibody was designed that has a markedly enhanced resistance to thermal inactivation and displays an unanticipated ≈30-fold improvement in affinity. Such supercharged antibodies should prove useful for assays in resource-limited settings and for developing reagents with improved shelf lives.
- Published
- 2012
- Full Text
- View/download PDF