@article{105733d72ff748349c6784fbdbe99b1a,
title = "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 (T m) 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 T m compared to the parental scFv and importantly, to retain full antigen binding activity after 2 hours of incubation at 70 °C. The crystal structure of one thermostabilized scFv variants was solved at 1.6 {\AA} and shown to be in close agreement with the RosettaAntibody model prediction. ",
keywords = "Rosetta, antibody engineering, scAb, scFv, thermostable antibodies",
author = "Jiwon Lee and Der, {Bryan S.} and Karamitros, {Christos S.} and Wenzong Li and Marshall, {Nicholas M.} and Lungu, {Oana I.} and Miklos, {Aleksandr E.} and Jianqing Xu and Kang, {Tae Hyun} and Lee, {Chang Han} and Bing Tan and Hughes, {Randall A.} and Jung, {Sang Taek} and Ippolito, {Gregory C.} and Gray, {Jeffrey J.} and Yan Zhang and Brian Kuhlman and George Georgiou and Ellington, {Andrew D.}",
note = "Funding Information: This work was supported by DARPA‐09‐69‐ATP‐FP‐005 (G.G., A.E., B.K., and J.G.), the Clayton Foundation (G.G.), NIH R01‐GM078221 (J.X. and J.G.), NIH R35‐GM131923 (B.K.), NIH R01‐125882 (Y.Z.), Welch Foundation F‐1654 (A.E.), and Welch Foundation F‐1778 (Y.Z.). Funding Information: This work was supported by DARPA-09-69-ATP-FP-005 (G.G., A.E., B.K., and J.G.), the Clayton Foundation (G.G.), NIH R01-GM078221 (J.X. and J.G.), NIH R35-GM131923 (B.K.), NIH R01-125882 (Y.Z.), Welch Foundation F-1654 (A.E.), and Welch Foundation F-1778 (Y.Z.). Publisher Copyright: {\textcopyright} 2019 American Institute of Chemical Engineers",
year = "2020",
month = mar,
day = "1",
doi = "10.1002/aic.16864",
language = "English (US)",
volume = "66",
journal = "AIChE Journal",
issn = "0001-1541",
publisher = "American Institute of Chemical Engineers",
number = "3",
}