Enhancing the Affinity of a Novel Selective scFv for Soluble ST2 through Computational Design
Issued Date
2025-01-01
Resource Type
ISSN
15499596
eISSN
1549960X
Scopus ID
2-s2.0-105005943336
Journal Title
Journal of Chemical Information and Modeling
Rights Holder(s)
SCOPUS
Bibliographic Citation
Journal of Chemical Information and Modeling (2025)
Suggested Citation
Matthies D.J., Novoa-Gundel P., Vásquez G., Dubois-Camacho K., De la fuente López M., Donoso B., Toledo-Stuardo K., Gutiérrez-González M., Landskron G., Valdebenito-Silva S., Sánchez O., Fierro A., Teimoori S., Chaicumpa W., Eugenin E., Zapata-Torres G., Molina M.C., Hermoso M.A. Enhancing the Affinity of a Novel Selective scFv for Soluble ST2 through Computational Design. Journal of Chemical Information and Modeling (2025). doi:10.1021/acs.jcim.4c02027 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/110449
Title
Enhancing the Affinity of a Novel Selective scFv for Soluble ST2 through Computational Design
Author's Affiliation
Universidad Finis Terrae
Pontificia Universidad Católica de Chile
The University of Texas Medical Branch at Galveston
Universidad de Chile
Faculty of Medicine Siriraj Hospital, Mahidol University
Universitair Medisch Centrum Groningen
Facultad de Medicina de la Universidad de Chile
Universidad de Concepcion
biotechrabbit GmbH
Pontificia Universidad Católica de Chile
The University of Texas Medical Branch at Galveston
Universidad de Chile
Faculty of Medicine Siriraj Hospital, Mahidol University
Universitair Medisch Centrum Groningen
Facultad de Medicina de la Universidad de Chile
Universidad de Concepcion
biotechrabbit GmbH
Corresponding Author(s)
Other Contributor(s)
Abstract
Suppression of Tumorigenicity 2 (ST2) is a member of the IL-1 receptor family, which includes transmembrane (ST2L) and soluble (sST2) isoforms. sST2 functions as a decoy receptor for Interleukin-33 (IL-33), thereby blocking the activation of the IL-33/ST2L signaling axis, which is essential for tissue repair and immune regulation. Clinical evidence indicates that elevated sST2 levels are associated with increased disease severity in conditions such as ulcerative colitis (UC), cardiovascular disease, and asthma. However, current antibodies cannot reliably distinguish between sST2 and its membrane-bound isoform ST2L, limiting their effectiveness for diagnostic and therapeutic use. To address this limitation, we developed an antibody that selectively targets sST2. Using a phage display library, we identified a single-chain variable fragment (scFv) with high specificity for a unique five amino acid sequence (SKECF) located at the C-terminus of sST2. Our parental scFv showed high selectivity for sST2 with minimal cross-reactivity to ST2L, as demonstrated by both flow cytometry and immunoprecipitation. Molecular simulations identified key binding residues, allowing the design of four scFv mutants, three of which displayed improved binding in surface plasmon resonance (SPR) analyses. The A183YL2 mutant exhibited a 3.4-fold increase in binding affinity, while G100WH3 demonstrated reduced binding due to unfavorable conformations. This study presents an anti-sST2 scFv with enhanced specificity and affinity, offering a promising tool for the diagnosis and treatment of inflammatory diseases, in which sST2 interferes with IL-33-mediated tissue repair.