Abstract
BackgroundAn affibody-displaying bio-nanocapsule (ZHER2-BNC) with a hepatocyte specificity derived from hepatitis B virus (HBV) was converted into an affibody, ZHER2, that recognizes HER2 receptors. This affibody was previously reported to be the result of the endocytosis-dependent specific uptake of proteins and siRNA into target cancer cells. To assist the endosomal escape of inclusions, a helper lipid with pH-sensitive fusogenic ability (1,2-dioleoyl-sn-glycero-3-phos phoethanolamine; DOPE) was conjugated with a ZHER2-BNC.FindingsIn this study, we displayed a pH-sensitive fusogenic GALA peptide on the surface of a particle in order to confer the ability of endosomal escape to a ZHER2-BNC. A GALA-displaying ZHER2-BNC purified from yeast uneventfully formed a particle structure. Furthermore, endosomal escape of the particle was facilitated after endocytic uptake and release of the inclusions to the cytoplasm without the cell toxicity.ConclusionThe genetic fusion of a GALA peptide to the virus-like particle confers the ability of endosomal escape.
Highlights
An affibody-displaying bio-nanocapsule (ZHER2-BNC) with a hepatocyte specificity derived from hepatitis B virus (HBV) was converted into an affibody, ZHER2, that recognizes Human epidermal growth factor receptor 2 (HER2) receptors
To accomplish endosomal escape, the fusion was designed to orient the GALA peptide in the direction of the outer surface of the ZHER2-BNC
Sample was subjected to SDS-Poly-acrylamide gel electrophoresis (PAGE) followed by immune blotting using anti-His6 antibody and anti-protein A antibody. (C) Size distribution using dynamic light scattering (DLS) analysis
Summary
An affibody-displaying bio-nanocapsule (ZHER2-BNC) with a hepatocyte specificity derived from hepatitis B virus (HBV) was converted into an affibody, ZHER2, that recognizes HER2 receptors. The ZHER2-BNC has succeeded in specific deliveries and function expressions of proteins or siRNA to the target cells by conjugating to a mixture of a specialized, cationic liposome (LP) with the ability for endosomal escape (1,2-dioleoyl-sn-glycero-3-phos phoethanolamine; DOPE) and an anionic LP [14,15]. To display a GALA peptide on the surface of ZHER2-BNC, it was genetically fused to a ZHER2-L protein.
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