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Structure-based design of supercharged, highly thermoresistant antibodies.

ABSTRACT: 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.

SUBMITTER: Miklos AE

PROVIDER: S-EPMC5583727 | biostudies-literature | 2012 Apr

REPOSITORIES: biostudies-literature

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Structure-based design of supercharged, highly thermoresistant antibodies.

Miklos Aleksandr E AE Kluwe Christien C Der Bryan S BS Pai Supriya S Sircar Aroop A Hughes Randall A RA Berrondo Monica M Xu Jianqing J Codrea Vlad V Buckley Patricia E PE Calm Alena M AM Welsh Heather S HS Warner Candice R CR Zacharko Melody A MA Carney James P JP Gray Jeffrey J JJ Georgiou George G Kuhlman Brian B Ellington Andrew D AD

Chemistry & biology 20120401 4

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. ...[more]

PMID: 22520751

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