Computer-based engineering of thermostabilized antibody fragments

Jiwon Lee, Bryan S. Der, Christos S. Karamitros, Wenzong Li, Nicholas M. Marshall, Oana I. Lungu, Aleksandr E. Miklos, Jianqing Xu, Tae Hyun Kang, Chang Han Lee, Bing Tan, Randall A. Hughes, Sang Taek Jung, Gregory C. Ippolito, Jeffrey J. Gray, Yan Zhang, Brian Kuhlman, George Georgiou, Andrew D. Ellington

Research output: Contribution to journalArticle

3 Scopus citations

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.

Original languageEnglish (US)
Article numbere16864
JournalAIChE Journal
Volume66
Issue number3
DOIs
StatePublished - Mar 1 2020

Keywords

  • Rosetta
  • antibody engineering
  • scAb
  • scFv
  • thermostable antibodies

ASJC Scopus subject areas

  • Biotechnology
  • Environmental Engineering
  • Chemical Engineering(all)

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