Počet záznamů: 1
Protein Binder (ProBi) as a New Class of Structurally Robust Non-Antibody Protein Scaffold for Directed Evolution
- 1.0542184 - BTÚ 2022 RIV CH eng J - Článek v odborném periodiku
Pham, Phuong Ngoc - Huličiak, Maroš - Biedermannová, Lada - Černý, Jiří - Charnavets, Tatsiana - Fuertes, Gustavo - Herynek, Štěpán - Kolářová, Lucie - Kolenko, Petr - Pavlíček, Jiří - Zahradník, Jiří - Mikulecký, Pavel - Schneider, Bohdan
Protein Binder (ProBi) as a New Class of Structurally Robust Non-Antibody Protein Scaffold for Directed Evolution.
Viruses. Roč. 13, č. 2 (2021), č. článku 190. E-ISSN 1999-4915
Grant CEP: GA ČR(CZ) GA16-20507S; GA MŠMT(CZ) LM2015043; GA MŠMT(CZ) LM2018127
Institucionální podpora: RVO:86652036
Klíčová slova: directed evolution * protein scaffold * protein engineering * computational saturation * ribosome display * interleukin-10
Obor OECD: Virology
Impakt faktor: 5.818, rok: 2021
Způsob publikování: Open access
https://www.mdpi.com/1999-4915/13/2/190
Engineered small non-antibody protein scaffolds are a promising alternative to antibodies and are especially attractive for use in protein therapeutics and diagnostics. The advantages include smaller size and a more robust, single-domain structural framework with a defined binding surface amenable to mutation. This calls for a more systematic approach in designing new scaffolds suitable for use in one or more methods of directed evolution. We hereby describe a process based on an analysis of protein structures from the Protein Data Bank and their experimental examination. The candidate protein scaffolds were subjected to a thorough screening including computational evaluation of the mutability, and experimental determination of their expression yield in E. coli, solubility, and thermostability. In the next step, we examined several variants of the candidate scaffolds including their wild types and alanine mutants. We proved the applicability of this systematic procedure by selecting a monomeric single-domain human protein with a fold different from previously known scaffolds. The newly developed scaffold, called ProBi (Protein Binder), contains two independently mutable surface patches. We demonstrated its functionality by training it as a binder against human interleukin-10, a medically important cytokine. The procedure yielded scaffold-related variants with nanomolar affinity.
Trvalý link: http://hdl.handle.net/11104/0319662
Počet záznamů: 1