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Molecular Biology

, Volume 51, Issue 6, pp 865–873 | Cite as

Bifunctional Toxin DARP-LoPE Based on the Her2-Specific Innovative Module of a Non-Immunoglobulin Scaffold as a Promising Agent for Theranostics

  • G. M. Proshkina
  • D. V. Kiseleva
  • O. N. Shilova
  • A. V. Ryabova
  • E. I. Shramova
  • O. A. Stremovskiy
  • S. M. Deyev
Current Trends in the Application of Monoclonal Antibodies Special Issue

Abstract

We have generated and characterized HER2-specific targeted toxin based on the low-immunogenic variant of Pseudomonas exotoxin A (LoPE), in which most of the human immunodominant B-cell epitopes have been inactivated. Nonimmunoglobulin DARPin-based HER2-specific protein was used as a targeting module for toxin delivery to the cellular target. Using confocal microscopy, it has been found that both domains in this hybrid toxin retained their functionality, i.e., the specific interaction with HER2 receptor, as well as the internalization and effective transport to ER typical of the wild-type Pseudomonas exotoxin A. The HER2-dependent cytotoxic effect correlated with receptor expression level at the cell surface, as shown in vitro using cell lines with different levels of HER2 expression. Due to the very high selective cytotoxicity against HER2-positive human tumor cells, as well as expected low immunogenicity, we believe that this new targeted toxin may be promising for future in vivo studies as a therapeutic agent for HER2-positive tumors.

Keywords

DARPin targeted therapy Pseudomonas exotoxin А HER2 

Abbreviations

DARPin

designed ankyrin repeat proteins

HER2

human epidermal growth factor receptor 2

FITC

fluorescein isothiocyanate

PE

Pseudomonas exotoxin

ER

endoplasmic reticulum.

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Copyright information

© Pleiades Publishing, Inc. 2017

Authors and Affiliations

  • G. M. Proshkina
    • 1
  • D. V. Kiseleva
    • 1
    • 2
  • O. N. Shilova
    • 1
  • A. V. Ryabova
    • 3
  • E. I. Shramova
    • 1
  • O. A. Stremovskiy
    • 1
  • S. M. Deyev
    • 1
    • 4
  1. 1.Shemyakin–Ovchinnikov Institute of Bioorganic ChemistryRussian Academy of SciencesMoscowRussia
  2. 2.Department of BiologyMoscow State UniversityMoscowRussia
  3. 3.Prokhorov General Physics InstituteRussian Academy of SciencesMoscowRussia
  4. 4.National Research Tomsk Polytechnic UniversityTomskRussia

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