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

Le résumé fourni par la source

The FEBS Journal FJ-22-0728 Combined in vitro and cell-based selection display method producing specific binders against IL-9 receptor in high yields Maroš Huličiak, Lada Biedermanová, Daniel Berdár, Štěpán Herynek, Lucie Kolářová, Jakub Tomala, Pavel Mikulecký, and Bohdan Schneider Institute of Biotechnology of the Czech Academy of Sciences, Průmyslová 595, 252 50 Vestec, Czech Republic Correspondence: Bohdan Schneider, Institute of Biotechnology of the Czech Academy of Sciences, BIOCEV, CZ-252 50 Vestec, Czech Republic, Tel: +420 728 303 566, e-mail: bohdan.schneider@ibt.cas.cz Abstract 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 signaling 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.

Ce résumé expose les affirmations des auteurs. BNTIC ne l’interprète pas comme une validation indépendante des résultats.