title: 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.
authors.affiliation: Center for Systems and Synthetic Biology, University of Texas at Austin, Austin, TX 78712, USA.
authors.forename: Aleksandr E
authors.initials: AE
authors.lastname: Miklos
authors.forename: Christien
authors.initials: C
authors.lastname: Kluwe
authors.forename: Bryan S
authors.initials: BS
authors.lastname: Der
authors.forename: Supriya
authors.initials: S
authors.lastname: Pai
authors.forename: Aroop
authors.initials: A
authors.lastname: Sircar
authors.forename: Randall A
authors.initials: RA
authors.lastname: Hughes
authors.forename: Monica
authors.initials: M
authors.lastname: Berrondo
authors.forename: Jianqing
authors.initials: J
authors.lastname: Xu
authors.forename: Vlad
authors.initials: V
authors.lastname: Codrea
authors.forename: Patricia E
authors.initials: PE
authors.lastname: Buckley
authors.forename: Alena M
authors.initials: AM
authors.lastname: Calm
authors.forename: Heather S
authors.initials: HS
authors.lastname: Welsh
authors.forename: Candice R
authors.initials: CR
authors.lastname: Warner
authors.forename: Melody A
authors.initials: MA
authors.lastname: Zacharko
authors.forename: James P
authors.initials: JP
authors.lastname: Carney
authors.forename: Jeffrey J
authors.initials: JJ
authors.lastname: Gray
authors.forename: George
authors.initials: G
authors.lastname: Georgiou
authors.forename: Brian
authors.initials: B
authors.lastname: Kuhlman
authors.forename: Andrew D
authors.initials: AD
authors.lastname: Ellington
mesh_terms: D006860:Hydrogen Bonding; D015202:Protein Engineering; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D057127:Single-Chain Antibodies / Q000737:chemistry* / Q000378:metabolism; D012984:Software; D013696:Temperature
chemical_list: D057127:Single-Chain Antibodies