Engineering a New IFN-ApoA-I Fusion Protein with Low Toxicity and Prolonged Action

Svetlana Miroshnichenko, Mariya Pykhtina, Anastasiia Kotliarova, Alexander Chepurnov, Anatoly Beklemishev

Research output: Contribution to journalArticlepeer-review

Abstract

Recombinant human interferon alpha-2b (rIFN) is widely used in antiviral and anticancer immunotherapy. However, the high efficiency of interferon therapy is accompanied by a number of side effects; this problem requires the design of a new class of interferon molecules with reduced cytotoxicity. In this work, IFN was modified via genetic engineering methods by merging it with the blood plasma protein apolipoprotein A-I in order to reduce acute toxicity and improve the pharmacokinetics of IFN. The chimeric protein was obtained via biosynthesis in the yeast P. pastoris. The yield of ryIFN-ApoA-I protein when cultivated on a shaker in flasks was 30 mg/L; protein purification was carried out using reverse-phase chromatography to a purity of 95–97%. The chimeric protein demonstrated complete preservation of the biological activity of IFN in the model of vesicular stomatitis virus and SARS-CoV-2. In addition, the chimeric form had reduced cytotoxicity towards Vero cells and increased cell viability under viral load conditions compared with commercial IFN-a2b preparations. Analysis of the pharmacokinetic profile of ryIFN-ApoA-I after a single subcutaneous injection in mice showed a 1.8-fold increased half-life of the chimeric protein compared with ryIFN.

Original languageEnglish
Article number8014
JournalMolecules
Volume28
Issue number24
DOIs
StatePublished - 1 Dec 2023
Externally publishedYes

Keywords

  • Pichia pastoris
  • SARS-CoV-2
  • Tween 20
  • antiviral activity
  • apolipoprotein A-I (ApoA-I)
  • apoptosis
  • cytotoxicity
  • fusion protein
  • prolonged half-life
  • yeast-derived IFNα2b (ryIFN)

ASJC Scopus subject areas

  • Drug Discovery
  • Analytical Chemistry
  • Chemistry (miscellaneous)
  • Molecular Medicine
  • Physical and Theoretical Chemistry
  • Pharmaceutical Science
  • Organic Chemistry

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