Combined in vitro and cell-based selection display method producing specific binders against IL-9 receptor in high yields

0569019 - BTÚ 2024 RIV GB eng J - Journal Article
Huličiak, Maroš - Biedermannová, Lada - Berdár, Daniel - Herynek, Štěpán - Kolářová, Lucie - Tomala, Jakub - Mikulecký, Pavel - Schneider, Bohdan
Combined in vitro and cell-based selection display method producing specific binders against IL-9 receptor in high yields.
FEBS Journal. Roč. 290, č. 11 (2023), s. 2993-3005. ISSN 1742-464X. E-ISSN 1742-4658
R&D Projects: GA MŠMT LX22NPO5102; GA MŠMT(CZ) LM2018127; GA MŠMT(CZ) EF18_046/0015974; GA ČR(CZ) GA20-13029S
Institutional support: RVO:86652036
Keywords : directed evolution * interleukin 9 receptor alpha * protein scaffolds * ribosome display * yeast display
OECD category: Biochemistry and molecular biology
Impact factor: 5.4, year: 2022
Method of publishing: Open access
https://febs.onlinelibrary.wiley.com/doi/10.1111/febs.16726

We combined cell-free ribosome display and cell-based yeast display selection to build specific protein binders to the extracellular domain of the human interleukin 9 receptor alpha (IL-9Rα). The target, IL-9Rα, is the receptor involved in the signalling pathway of IL-9, a pro-inflammatory cytokine medically important for its involvement in respiratory diseases. The successive use of modified protocols of ribosome and yeast displays allowed us to combine their strengths—the virtually infinite selection power of ribosome display and the production of (mostly) properly folded and soluble proteins in yeast display. The described experimental protocol is optimized to produce binders highly specific to the target, including selectivity to common proteins such as BSA, and proteins potentially competing for the binder such as receptors of other cytokines. The binders were trained from DNA libraries of two protein scaffolds called 57aBi and 57bBi developed in our laboratory. We show that the described unconventional combination of ribosome and yeast displays is effective in developing selective small protein binders to the medically relevant molecular target.
Permanent Link: https://hdl.handle.net/11104/0340309