Archivum Immunologiae et Therapiae Experimentalis

, Volume 61, Issue 1, pp 75–83

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

  • Alireza Shoae-Hassani
  • Seyed Abdolreza Mortazavi-Tabatabaei
  • Shiva Sharif
  • Shabnam Madadi
  • Hamidreza Rezaei-Khaligh
  • Javad Verdi
Original Article

DOI: 10.1007/s00005-012-0206-x

Cite this article as:
Shoae-Hassani, A., Mortazavi-Tabatabaei, S.A., Sharif, S. et al. Arch. Immunol. Ther. Exp. (2013) 61: 75. doi:10.1007/s00005-012-0206-x
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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.

Keywords

Nanomedicine Nanobody Nanobioparticle Recombinant 

Copyright information

© L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012

Authors and Affiliations

  • Alireza Shoae-Hassani
    • 1
  • Seyed Abdolreza Mortazavi-Tabatabaei
    • 2
    • 1
  • Shiva Sharif
    • 3
    • 1
  • Shabnam Madadi
    • 4
  • Hamidreza Rezaei-Khaligh
    • 1
  • Javad Verdi
    • 1
    • 3
    • 5
  1. 1.Research Center for Science and Technology in Medicine (RCSTIM)Tehran University of Medical Sciences (TUMS)TehranIran
  2. 2.Proteomics Research CenterShahid Beheshti University of Medical SciencesTehranIran
  3. 3.Department of Tissue EngineeringSchool of Advanced Technologies in Medicine, Tehran University of Medical SciencesTehranIran
  4. 4.Cardiac Imaging Center, Shaheed Rajai Cardiovascular CenterTehran University of Medical SciencesTehranIran
  5. 5.Department of Applied Cell SciencesSchool of Advanced Technologies in Medicine, Tehran University of Medical SciencesTehranIran

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