Computer‐based engineering of thermostabilized antibody fragments
Abstract
We used the molecular modeling program Rosetta to identify clusters of amino acid substitutions in antibody fragments (scFvs and scAbs) that improve global protein stability and resistance to thermal deactivation. Using this methodology, we increased the melting temperature (Tm) and resistance to heat treatment of an antibody fragment that binds to the Clostridium botulinum hemagglutinin protein (anti‐HA33). Two designed antibody fragment variants with two amino acid replacement clusters, designed to stabilize local regions, were shown to have both higher Tm compared to the parental scFv and importantly to retain full antigen binding activity after 2 hr of incubation at 70°C. The crystal structure of one thermostabilized scFv variants was solved at 1.6 Å and shown to be in close agreement with the RosettaAntibody model prediction.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Nov 29, 2019
- Source ID
- 10.1002/aic.16864
Entities
People
- Aleksandr E. Miklos
- Andrew D Ellington
- Bing Tan
- Brian Kuhlman
- Bryan S. Der
- Chang‐han Lee
- Christos S. Karamitros
- George Georgiou
- Gregory C Ippolito
- Jeffrey J. Gray
- Jianqing Xu
- Jiwon Lee
- Nicholas M. Marshall
- Oana I. Lungu
- Randall A. Hughes
- Sang Taek Jung
- Tae Hyun Kang
- Wenzong Li
- Yan Zhang
Organizations
- Dartmouth College
- Defense Advanced Research Projects Agency
- Johns Hopkins University
- Kookmin University
- Korea University
- National Institutes of Health
- Robert A. Welch Foundation
- United States Army Research Laboratory
- University of North Carolina
- University of Texas at Austin