Recombinant λ Bacteriophage Displaying Nanobody towards Third Domain of HER-2 Epitope Inhibits Proliferation of Breast Carcinoma SKBR-3 Cell Line

Abstract

Phage display of many nanobodies via filamentous phage in combination with helper phage has been reported by many scientists. The aim of this study was to produce lambda (λ) bacteriophage displaying high-affinity nanobody against HER-2 expressing breast carcinoma cells. Bacteriophage λ is a temperate phage with inherent biological safety in mammalian cells. Here we report the construction of a recombinant λ phage that efficiently expresses specific nanobody towards third domain of HER-2 target on SKBR-3 and MCF-7 cell lines in vitro. We constructed recombinant λ phage particles containing a mammalian expression cassette, C-Myc tagged, encoding VHH gene of camelid anti HER-2 third domain epitope using λ ZAP-cytomegalic virus (CMV) vector. The SKBR-3, MCF-7 and human endometrial stem cells were treated by the nanobody displayed recombinant λ phage. The cell growth inhibition assay was performed by MTT Cell Viability Assay Kit. After the fourth round of biopanning there was a significant enrichment in the phage specifically binding to the antigen. The ratio of targeted phage increased approximately 1,000-fold in the fifth round. The nanobody expressed by λ ZAP-CMV-VHH phagemid cloned in λ bioparticles significantly inhibited the proliferation of HER-2 positive SKBR-3 and MCF-7 cells. Recombinant bacteriophage λ ZAP-CMV-VHH-cDNA could be used efficiently for construction of nanobodies to mortify HER-2 positive breast carcinoma cells as a nanomedical therapeutic.

Neurogenesis and Increase in Differentiated Neural Cell Survival via Phosphorylation of Akt1 after Fluoxetine Treatment of Stem Cells

“…Total cell extracts containing equal amounts of protein in sample buffer were subjected to WB analysis as described previously [13]. Briefly, the samples were boiled for 5 min and then separated by 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE).…”

λ Phage Nanobioparticle Expressing Apoptin Efficiently Suppress Human Breast Carcinoma Tumor Growth In Vivo

“…As described, the filamentous phages have been used for gene delivery vectors in some cases but there are few reports about using lambda phages for this purpose. Recently we showed that lambda phage could express nanobody against her-2 positive breast carcinoma cells [34] . In this work the human breast carcinoma was stopped efficiently by λ NBPs but there was no apoptosis in normal cells (BMSCs).…”

Preparation of a transferrin-targeted M13-based gene nanocarrier and evaluation of its efficacy for gene delivery and expression in eukaryote cells

“…Chemical coupling has been applied as a straightforward strategy for linking targeting molecules and/or drugs with nanoparticles/nanobioparticles. In particular, this strategy has achieved some insights in imaging (Singh et al, 2006;Li et al, 2010;Huang et al, 2011) and drug/gene delivery (Stephanopoulos et al, 2010;Khalaj-Kondori et al, 2011;Shoae-Hassani et al, 2013a, 2013b using targeted bacteriophage particles.…”