Structure-Based Design of Supercharged, Highly Thermoresistant Antibodies

被引：120

作者：

Miklos, Aleksandr E. ^{[1
,2
,3
]}

Kluwe, Christien ^{[1
,2
]}

Der, Bryan S. ^{[4
]}

Pai, Supriya ^{[2
]}

Sircar, Aroop ^{[5
]}

Hughes, Randall A. ^{[1
,2
,3
]}

Berrondo, Monica ^{[5
]}

Xu, Jianqing ^{[5
]}

Codrea, Vlad ^{[1
]}

Buckley, Patricia E. ^{[6
]}

Calm, Alena M. ^{[6
]}

Welsh, Heather S. ^{[6
]}

Warner, Candice R. ^{[7
]}

Zacharko, Melody A. ^{[7
]}

Carney, James P. ^{[6
]}

Gray, Jeffrey J. ^{[5
]}

Georgiou, George ^{[2
]}

Kuhlman, Brian ^{[4
]}

Ellington, Andrew D. ^{[1
,2
,3
]}

机构：

[1] Univ Texas Austin, Ctr Syst & Synthet Biol, Austin, TX 78712 USA

[2] Univ Texas Austin, Inst Cellular & Mol Biol, Austin, TX 78712 USA

[3] Univ Texas Austin, Appl Res Labs, Austin, TX 78712 USA

[4] Univ N Carolina, Dept Biochem & Biophys, Chapel Hill, NC 27599 USA

[5] Johns Hopkins Univ, Dept Chem & Biomol Engn, Baltimore, MD 21218 USA

[6] USA, Biosci Div, Edgewood Biol Chem Res Ctr, Aberdeen Proving Ground, MD 21010 USA

[7] Excet Inc, Springfield, VA 22151 USA

来源：

CHEMISTRY & BIOLOGY | 2012年 / 19卷 / 04期

关键词：

PROTEIN INTERACTIONS; PHAGE DISPLAY; AGGREGATION; STABILITY; BINDING;

D O I：

10.1016/j.chembiol.2012.01.018

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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 approximate to 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.

引用

页码：449 / 455

页数：7

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