Miklos Aleksandr E, Kluwe Christien, Der Bryan S, Pai Supriya, Sircar Aroop, Hughes Randall A, Berrondo Monica, Xu Jianqing, Codrea Vlad, Buckley Patricia E, Calm Alena M, Welsh Heather S, Warner Candice R, Zacharko Melody A, Carney James P, Gray Jeffrey J, Georgiou George, Kuhlman Brian, Ellington Andrew D
Center for Systems and Synthetic Biology, University of Texas at Austin, Austin, TX 78712, USA.
Chem Biol. 2012 Apr 20;19(4):449-55. doi: 10.1016/j.chembiol.2012.01.018.
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.
将表面残基突变为带电荷的氨基酸可增加对聚集的抵抗力，并能实现可逆解折叠。我们开发了一种使用Rosetta计算设计软件包的方案，该方案在考虑每个突变的能量影响的同时对蛋白质进行“超荷化”。利用同源模型，设计了一种单链可变片段抗体，它对热失活具有显著增强的抵抗力，并且亲和力有意外的约30倍的提高。这种超荷化抗体在资源有限的环境中的检测以及开发保质期更长的试剂方面应该会很有用。
