title: 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.
authors.affiliation: Research Center for Science and Technology in Medicine (RCSTIM), Tehran University of Medical Sciences (TUMS), Tehran, Iran.
authors.forename: Alireza
authors.initials: A
authors.lastname: Shoae-Hassani
authors.forename: Seyed Abdolreza
authors.initials: SA
authors.lastname: Mortazavi-Tabatabaei
authors.forename: Shiva
authors.initials: S
authors.lastname: Sharif
authors.forename: Shabnam
authors.initials: S
authors.lastname: Madadi
authors.forename: Hamidreza
authors.initials: H
authors.lastname: Rezaei-Khaligh
authors.forename: Javad
authors.initials: J
authors.lastname: Verdi
mesh_terms: D053687:Adult Stem Cells / Q000187:drug effects; D000818:Animals; D000915:Antibody Affinity; D010582:Bacteriophage lambda; D001943:Breast Neoplasms / Q000276:immunology* / Q000628:therapy; D002161:Camelids, New World; D002277:Carcinoma / Q000276:immunology* / Q000628:therapy; D049109:Cell Proliferation / Q000187:drug effects; D061505:Cell Surface Display Techniques / Q000379:methods*; D003587:Cytomegalovirus / Q000235:genetics; D018985:Epitopes, B-Lymphocyte / Q000276:immunology; D016259:Genes, myc / Q000235:genetics; D005818:Genetic Engineering; D006801:Humans; D061986:MCF-7 Cells; D017434:Protein Structure, Tertiary / Q000235:genetics; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics / Q000276:immunology; D061905:Single-Domain Antibodies / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology*
chemical_list: D018985:Epitopes, B-Lymphocyte; D018719:Erb-b2 Receptor Tyrosine Kinases; D061905:Single-Domain Antibodies; C508053:ERBB2 protein, human