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

doi:10.1002/aic.16864

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

Whiting School of Engineering

Research output: Contribution to journal › Article › peer-review

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 (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 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 language	English (US)
Article number	e16864
Journal	AIChE Journal
Volume	66
Issue number	3
DOIs	https://doi.org/10.1002/aic.16864
State	Published - Mar 1 2020

Keywords

Rosetta
antibody engineering
scAb
scFv
thermostable antibodies

ASJC Scopus subject areas

Biotechnology
Environmental Engineering
General Chemical Engineering

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10.1002/aic.16864

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Lee, J., Der, B. S., Karamitros, C. S., Li, W., Marshall, N. M., Lungu, O. I., Miklos, A. E., Xu, J., Kang, T. H., Lee, C. H., Tan, B., Hughes, R. A., Jung, S. T., Ippolito, G. C., Gray, J. J., Zhang, Y., Kuhlman, B., Georgiou, G., & Ellington, A. D. (2020). Computer-based engineering of thermostabilized antibody fragments. AIChE Journal, 66(3), Article e16864. https://doi.org/10.1002/aic.16864

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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 (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 {\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 = "Publisher Copyright: {\textcopyright} 2019 American Institute of Chemical Engineers",

year = "2020",

month = mar,

day = "1",

doi = "10.1002/aic.16864",

language = "English (US)",

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AU - Lee, Jiwon

AU - Der, Bryan S.

AU - Karamitros, Christos S.

AU - Li, Wenzong

AU - Marshall, Nicholas M.

AU - Lungu, Oana I.

AU - Miklos, Aleksandr E.

AU - Xu, Jianqing

AU - Kang, Tae Hyun

AU - Lee, Chang Han

AU - Tan, Bing

AU - Hughes, Randall A.

AU - Jung, Sang Taek

AU - Ippolito, Gregory C.

AU - Gray, Jeffrey J.

AU - Zhang, Yan

AU - Kuhlman, Brian

AU - Georgiou, George

AU - Ellington, Andrew D.

PY - 2020/3/1

Y1 - 2020/3/1

N2 - 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.

AB - 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.

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Computer-based engineering of thermostabilized antibody fragments

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Keywords

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