Construction of Artificial TNF-Binding Proteins Based on the 10th Human Fibronectin Type III Domain Using Bacterial Display
- Authors: Shingarova L.N.1, Petrovskaya L.E.1, Zlobinov A.V.1,2, Gapizov S.S.1,2, Kryukova E.A.1, Birikh K.R.1, Boldyreva E.F.1, Yakimov S.A.1, Dolgikh D.A.1,2, Kirpichnikov M.P.1,2
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Affiliations:
- Shemyakin−Ovchinnikov Institute of Bioorganic Chemistry
- Lomonosov Moscow State University
- Issue: Vol 83, No 6 (2018)
- Pages: 708-716
- Section: Article
- URL: https://journals.rcsi.science/0006-2979/article/view/151673
- DOI: https://doi.org/10.1134/S0006297918060081
- ID: 151673
Cite item
Abstract
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.
About the authors
L. N. Shingarova
Shemyakin−Ovchinnikov Institute of Bioorganic Chemistry
Author for correspondence.
Email: lshing@ibch.ru
Russian Federation, Moscow, 117997
L. E. Petrovskaya
Shemyakin−Ovchinnikov Institute of Bioorganic Chemistry
Email: lshing@ibch.ru
Russian Federation, Moscow, 117997
A. V. Zlobinov
Shemyakin−Ovchinnikov Institute of Bioorganic Chemistry; Lomonosov Moscow State University
Email: lshing@ibch.ru
Russian Federation, Moscow, 117997; Moscow, 119234
S. Sh. Gapizov
Shemyakin−Ovchinnikov Institute of Bioorganic Chemistry; Lomonosov Moscow State University
Email: lshing@ibch.ru
Russian Federation, Moscow, 117997; Moscow, 119234
E. A. Kryukova
Shemyakin−Ovchinnikov Institute of Bioorganic Chemistry
Email: lshing@ibch.ru
Russian Federation, Moscow, 117997
K. R. Birikh
Shemyakin−Ovchinnikov Institute of Bioorganic Chemistry
Email: lshing@ibch.ru
Russian Federation, Moscow, 117997
E. F. Boldyreva
Shemyakin−Ovchinnikov Institute of Bioorganic Chemistry
Email: lshing@ibch.ru
Russian Federation, Moscow, 117997
S. A. Yakimov
Shemyakin−Ovchinnikov Institute of Bioorganic Chemistry
Email: lshing@ibch.ru
Russian Federation, Moscow, 117997
D. A. Dolgikh
Shemyakin−Ovchinnikov Institute of Bioorganic Chemistry; Lomonosov Moscow State University
Email: lshing@ibch.ru
Russian Federation, Moscow, 117997; Moscow, 119234
M. P. Kirpichnikov
Shemyakin−Ovchinnikov Institute of Bioorganic Chemistry; Lomonosov Moscow State University
Email: lshing@ibch.ru
Russian Federation, Moscow, 117997; Moscow, 119234
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