Selective staining and eradication of cancer cells by protein-carrying DARPin-functionalized liposomes

Sergey Deyev, Galina Proshkina, Olga Baryshnikova, Anastasiya Ryabova, Gavriel Avishai, Liat Katrivas, Clelia Giannini, Yael Levi-Kalisman, Alexander Kotlyar

Research output: Contribution to journalArticlepeer-review

6 Citations (Scopus)

Abstract

Since their discovery, liposomes have been widely employed in biomedical research. These nano-size spherical vesicles consisting one or few phospholipid bilayers surrounding an aqueous core are capable of carrying a wide variety of bioactive compounds, including drugs, peptides, nucleic acids, proteins and others. Despite considerable success achieved in synthesis of liposome constructs containing bioactive compounds, preparation of ligand-targeted liposomes comprising large quantities of encapsulated proteins that are capable of affecting pathological cells still remains a big challenge. Here we described a novel method for preparation of small (80–90 nm in diameter) unilamellar liposomes containing very large quantities (thousands of protein molecules per liposome) of heme-containing cytochrome c, highly fluorescent mCherry and highly toxic PE40 (Pseudomonas aeruginosa Exotoxin A domain). Efficient encapsulation of the proteins was achieved through electrostatic interaction between positively charged proteins (at pH lower than pI) and negatively charged liposome membrane. The proteoliposomes containing large quantities of mCherry or PE40 and functionalized with designed ankyrin repeat protein (DARPin)_9-29, which targets human epidermal growth factor receptor 2 (HER2) were shown to specifically stain and kill in sub-nanomolar concentrations HER2-positive cells, overexpressing HER2, respectively. Specific staining and eradication of the receptor-positive cells demonstrated here makes the DARPin-functionalized liposomes carrying large quantities of fluorescent and/or toxic proteins a promising candidate for tumor detection and therapy.

Original languageEnglish
Pages (from-to)296-305
Number of pages10
JournalEuropean Journal of Pharmaceutics and Biopharmaceutics
Volume130
DOIs
Publication statusPublished - Sep 2018
Externally publishedYes

Keywords

  • Apoptosis
  • Cancer
  • Cryo-TEM
  • DARPin
  • Exotoxin A
  • Liposomes
  • mCherry

ASJC Scopus subject areas

  • Biotechnology
  • Pharmaceutical Science

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