Biochemistry (Moscow)

, Volume 84, Issue 6, pp 627–636 | Cite as

A New Efficient Method for Production of Recombinant Antitumor Cytokine TRAIL and Its Receptor-Selective Variant DR5-B

  • A. V. Yagolovich
  • A. A. Artykov
  • D. A. Dolgikh
  • M. P. Kirpichnikov
  • M. E. GasparianEmail author


The cytokine TRAIL induces apoptosis in tumor cells of various origin without affecting normal cells. Clinical trials of TRAIL-receptor (DR4 and DR5) agonists (recombinant TRAIL or death receptors antibodies) have largely failed because most human tumors were resistant to them. Currently, a second generation of agents targeted at TRAIL-R with increased efficiency has been developed. To this end, we have developed DR5-B, a variant of TRAIL selectively interacting with DR5. We have developed a new efficient method for production of TRAIL and DR5-B using expression of these proteins in Escherichia coli strain SHuffle B. The proteins were isolated from the cytoplasmic fraction of cells and purified to a high degree of homogeneity using metal-affinity and ion-exchange chromatography. The protein yield was 211 and 173 mg from one liter of cell culture for DR5-B and TRAIL, respectively, which significantly exceeded the results obtained by other methods. DR5-B killed tumor cells of different origin more efficiently and rapidly compared with TRAIL. The resulting preparations can be used for the study of TRAIL signaling pathways and in preclinical and clinical trials as antitumor agents.


cytokine TRAIL mutant variant DR5-B strain E. coli SHuffle B cancer therapy 



death domain


death receptor


human foreskin fibroblasts




3-[4,5-dimethylthiazol-2]-2,5-diphenyl tetrazolium bromide




TNF-related apoptosis-inducing ligand


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Copyright information

© Pleiades Publishing, Ltd. 2019

Authors and Affiliations

  • A. V. Yagolovich
    • 1
    • 2
  • A. A. Artykov
    • 1
    • 2
  • D. A. Dolgikh
    • 1
    • 2
  • M. P. Kirpichnikov
    • 1
    • 2
  • M. E. Gasparian
    • 1
    Email author
  1. 1.Shemyakin and Ovchinnikov Institute of Bioorganic ChemistryRussian Academy of SciencesMoscowRussia
  2. 2.Faculty of BiologyLomonosov Moscow State UniversityMoscowRussia

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