Biochemistry (Moscow)

, Volume 83, Issue 6, pp 708–716 | Cite as

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

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


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.


tumor necrosis factor TNF-binding proteins 10th human fibronectin type III domain bacterial display autotransporter from Psychrobacter cryohalolentis K5T 



alternative scaffold protein




biotinylated TNF


complementarity-determining region


10th human fibronectin type III domain


isopropyl β-D-1-thiogalactopyranoside


3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide


phosphate buffered saline


phenyl-methylsulfonyl fluoride




tumor necrosis factor


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Copyright information

© Pleiades Publishing, Ltd. 2018

Authors and Affiliations

  • L. N. Shingarova
    • 1
  • L. E. Petrovskaya
    • 1
  • A. V. Zlobinov
    • 1
    • 2
  • S. Sh. Gapizov
    • 1
    • 2
  • E. A. Kryukova
    • 1
  • K. R. Birikh
    • 1
  • E. F. Boldyreva
    • 1
  • S. A. Yakimov
    • 1
  • D. A. Dolgikh
    • 1
    • 2
  • M. P. Kirpichnikov
    • 1
    • 2
  1. 1.Shemyakin−Ovchinnikov Institute of Bioorganic ChemistryRussian Academy of SciencesMoscowRussia
  2. 2.Lomonosov Moscow State UniversityFaculty of BiologyMoscowRussia

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