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

Huličiak, Maroš; Biedermannová, Lada; Berdár, Daniel; Herynek, Štěpán; Kolářová, Lucie; Tomala, Jakub; Mikulecký, Pavel; Schneider, Bohdan

doi:https://dx.doi.org/10.1111/febs.16726

Number of the records: 1

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

1.

SYSNO ASEP	0569019
Document Type	J - Journal Article
R&D Document Type	Journal Article
Subsidiary J	Článek ve WOS
Title	Combined in vitro and cell-based selection display method producing specific binders against IL-9 receptor in high yields
Author(s)	Huličiak, Maroš (BTO-N) Biedermannová, Lada (BTO-N)_{RID, ORCID} Berdár, Daniel (BTO-N) Herynek, Štěpán (BTO-N) Kolářová, Lucie (BTO-N) Tomala, Jakub (BTO-N) Mikulecký, Pavel (BTO-N)_RID Schneider, Bohdan (BTO-N)_{RID, ORCID}
Number of authors	8
Source Title	FEBS Journal - ISSN 1742-464X Roč. 290, č. 11 (2023), s. 2993-3005
Number of pages	13 s.
Language	eng - English
Country	GB - United Kingdom
Keywords	directed evolution ; interleukin 9 receptor alpha ; protein scaffolds ; ribosome display ; yeast display
Subject RIV	EB - Genetics ; Molecular Biology
OECD category	Biochemistry and molecular biology
R&D Projects	LX22NPO5102 GA MŠMT - Ministry of Education, Youth and Sports (MEYS)
	LM2018127 GA MŠMT - Ministry of Education, Youth and Sports (MEYS)
	EF18_046/0015974 GA MŠMT - Ministry of Education, Youth and Sports (MEYS)
	GA20-13029S GA ČR - Czech Science Foundation (CSF)
Method of publishing	Open access
Institutional support	BTO-N - RVO:86652036
UT WOS	000919058700001
EID SCOPUS	85147001025
DOI	10.1111/febs.16726
Annotation	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.
Workplace	Institute of Biotechnology
Contact	Monika Kopřivová, Monika.Koprivova@ibt.cas.cz, Tel.: 325 873 700
Year of Publishing	2024
Electronic address	https://febs.onlinelibrary.wiley.com/doi/10.1111/febs.16726

Number of the records: 1