Construction of Artificial TNF-Binding Proteins Based on the 10th Human Fibronectin Type III Domain Using Bacterial Display
- Autores: Shingarova L.1, Petrovskaya L.1, Zlobinov A.1,2, Gapizov S.1,2, Kryukova E.1, Birikh K.1, Boldyreva E.1, Yakimov S.1, Dolgikh D.1,2, Kirpichnikov M.1,2
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Afiliações:
- Shemyakin−Ovchinnikov Institute of Bioorganic Chemistry
- Lomonosov Moscow State University
- Edição: Volume 83, Nº 6 (2018)
- Páginas: 708-716
- Seção: Article
- URL: https://journals.rcsi.science/0006-2979/article/view/151673
- DOI: https://doi.org/10.1134/S0006297918060081
- ID: 151673
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Resumo
Construction of antibody mimetics on the base of alternative scaffold proteins is a promising strategy for obtaining new products for medicine and biotechnology. The aim of our work was to optimize the cell display system for the 10th human fibronectin type III domain (10Fn3) scaffold protein based on the AT877 autotransporter from Psychrobacter cryohalolentis K5T and to construct new artificial TNF-binding proteins. We obtained a 10Fn3 gene combinatorial library and screened it using the bacterial display method. After expression of the selected 10Fn3 variants in Escherichia coli cells and analysis of their TNF-binding activity, we identified proteins that display high affinity for TNF and characterized their properties.
Sobre autores
L. Shingarova
Shemyakin−Ovchinnikov Institute of Bioorganic Chemistry
Autor responsável pela correspondência
Email: lshing@ibch.ru
Rússia, Moscow, 117997
L. Petrovskaya
Shemyakin−Ovchinnikov Institute of Bioorganic Chemistry
Email: lshing@ibch.ru
Rússia, Moscow, 117997
A. Zlobinov
Shemyakin−Ovchinnikov Institute of Bioorganic Chemistry; Lomonosov Moscow State University
Email: lshing@ibch.ru
Rússia, Moscow, 117997; Moscow, 119234
S. Gapizov
Shemyakin−Ovchinnikov Institute of Bioorganic Chemistry; Lomonosov Moscow State University
Email: lshing@ibch.ru
Rússia, Moscow, 117997; Moscow, 119234
E. Kryukova
Shemyakin−Ovchinnikov Institute of Bioorganic Chemistry
Email: lshing@ibch.ru
Rússia, Moscow, 117997
K. Birikh
Shemyakin−Ovchinnikov Institute of Bioorganic Chemistry
Email: lshing@ibch.ru
Rússia, Moscow, 117997
E. Boldyreva
Shemyakin−Ovchinnikov Institute of Bioorganic Chemistry
Email: lshing@ibch.ru
Rússia, Moscow, 117997
S. Yakimov
Shemyakin−Ovchinnikov Institute of Bioorganic Chemistry
Email: lshing@ibch.ru
Rússia, Moscow, 117997
D. Dolgikh
Shemyakin−Ovchinnikov Institute of Bioorganic Chemistry; Lomonosov Moscow State University
Email: lshing@ibch.ru
Rússia, Moscow, 117997; Moscow, 119234
M. Kirpichnikov
Shemyakin−Ovchinnikov Institute of Bioorganic Chemistry; Lomonosov Moscow State University
Email: lshing@ibch.ru
Rússia, Moscow, 117997; Moscow, 119234