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Construction of Artificial TNF-Binding Proteins Based on the 10th Human Fibronectin Type III Domain Using Bacterial Display

by Kirpichnikov, M P , Zlobinov, A V , Kryukova, E A , Birikh, K R , Dolgikh, D A , Boldyreva, E F , Petrovskaya, L E , Yakimov, S A , S Sh Gapizov , Shingarova, L N

in Antibodies / Antigens / Binding / Biochemistry / Biotechnology / Cloning / Combinatorial analysis / Display devices / E coli / Efficiency / Fibronectin / Gene amplification / Proteins / Scaffolds / TNF inhibitors / Tumor necrosis factor-TNF

2018

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Construction of Artificial TNF-Binding Proteins Based on the 10th Human Fibronectin Type III Domain Using Bacterial Display

by Kirpichnikov, M P , Zlobinov, A V , Kryukova, E A , Birikh, K R , Dolgikh, D A , Boldyreva, E F , Petrovskaya, L E , Yakimov, S A , S Sh Gapizov , Shingarova, L N

2018

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Construction of Artificial TNF-Binding Proteins Based on the 10th Human Fibronectin Type III Domain Using Bacterial Display

by Kirpichnikov, M P , Zlobinov, A V , Kryukova, E A , Birikh, K R , Dolgikh, D A , Boldyreva, E F , Petrovskaya, L E , Yakimov, S A , S Sh Gapizov , Shingarova, L N

2018

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Journal Article

Construction of Artificial TNF-Binding Proteins Based on the 10th Human Fibronectin Type III Domain Using Bacterial Display

Kirpichnikov, M P,

Zlobinov, A V,

Kryukova, E A,

Birikh, K R,

Dolgikh, D A,

Boldyreva, E F,

Petrovskaya, L E,

Yakimov, S A,

S Sh Gapizov,

Shingarova, L N

2018

Overview

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.

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Publisher

Springer Nature B.V

Subject

/ Binding

/ Cloning

/ Combinatorial analysis