Towards canine immunotherapy models: Monoclonal antibodies with redox regulated epitopes targeting TIM3 attenuate Galectin-9 binding
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61389030%3A_____%2F26%3A00646729" target="_blank" >RIV/61389030:_____/26:00646729 - isvavai.cz</a>
Result on the web
<a href="https://doi.org/10.1016/j.ab.2025.116033" target="_blank" >https://doi.org/10.1016/j.ab.2025.116033</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1016/j.ab.2025.116033" target="_blank" >10.1016/j.ab.2025.116033</a>
Alternative languages
Result language
angličtina
Original language name
Towards canine immunotherapy models: Monoclonal antibodies with redox regulated epitopes targeting TIM3 attenuate Galectin-9 binding
Original language description
The TIM3 receptor acts as an immune checkpoint protein. Canine cancers exhibit higher pan-cancer penetrance of TIM3 compared to PD1, highlighting the potential of TIM3 as a compelling target in comparative immuno-oncology. We have used a highly diverse naïve canine scFv phage library to isolate antibodies to canine TIM3. Alternating rounds of biopanning were performed using either Fc or GST tagged canine TIM3 synthesized in mammalian cells. scFv sequences were identified using colony screening and next generation deep sequencing (NGS). The NGS protocol identified lower abundant frequency clones, demonstrating the enhanced depth of repertoire discovery possible using sequence-tag based tracing. Three representative scFv were expressed as mouse-canine chimeric scFv-Fc fusions or as full-length chimeric IgG. These antibodies all bound to the TIM3 receptor using either ELISA or immunoblotting. All the antibodies displayed sensitivity to reducing agents, which indicates the existence of disulfide-stabilized conformational epitopes. Epitope mapping using pepscan libraries suggested that the antibodies recognize a shared structural motif within the IgV domain of TIM3, within β-sheets fixed by disulfide bonds which would form the conformational epitope. Such conformational epitopes might be functional because they overlap with ligand-binding interfaces. Consistent with this, the antibodies attenuated TIM3 binding to Galectin-9. These data affirm that naïve canine scFv antibody libraries can yield self-antigen reactive antibodies to immune blockade receptor antigens. The data also emphasize the value in using native, folded, mammalian expressed receptor antigens to increase the probability of acquiring conformationally sensitive antibodies with potential therapeutic applications in veterinary and human medicine.
Czech name
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Czech description
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Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
10609 - Biochemical research methods
Result continuities
Project
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Continuities
I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace
Others
Publication year
2026
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Analytical Biochemistry
ISSN
0003-2697
e-ISSN
1096-0309
Volume of the periodical
711
Issue of the periodical within the volume
APR
Country of publishing house
NL - THE KINGDOM OF THE NETHERLANDS
Number of pages
21
Pages from-to
116033
UT code for WoS article
001665100000001
EID of the result in the Scopus database
2-s2.0-105026986566