title: Generation of an enriched pool of DNA aptamers for an HER2-overexpressing cell line selected by Cell SELEX.
abstract: Overexpression of human epidermal growth factor receptor 2 (HER2) occurs in a large percentage of breast cancers. Monoclonal antibodies targeting HER2 are vastly used for both diagnostic and therapeutic aims. However, identifying a new molecular probe against HER2 with improved diagnostic and therapeutic features is of great importance. In this report, we have applied the cell systematic evolution of ligands by exponential enrichment (SELEX) strategy for 16 selection rounds to generate an enriched pool of aptamers that specifically recognize the HER2 positive cell line. During the Cell SELEX procedure, a human HER2-overexpressing breast cancer cell line and a human HER2 negative breast cancer cell line were used. Our results reveal that polymerase chain reaction (PCR) amplification of random DNA libraries and the selected single-stranded DNA pool in different Cell SELEX rounds are different from what we expect from PCR amplification of homologous DNA. Our results also confirmed previous studies describing positive HER2 status of SK-BR3 and the absence of the HER2 expression in the MDA-MB468. We also developed a new method, Cell enzyme-linked assay, to monitor the enrichment of aptamers in a given round of Cell SELEX. This method would also be useful in other experiments using live cell enzyme-linked immunosorbent assay on adherent cells.
authors.affiliation: Department of Pathobiology, Ferdowsi University of Mashhad, Mashhad, Iran.
authors.forename: Kazem
authors.initials: K
authors.lastname: Dastjerdi
authors.forename: Gholamreza Hashemi
authors.initials: GH
authors.lastname: Tabar
authors.forename: Hesam
authors.initials: H
authors.lastname: Dehghani
authors.forename: Alireza
authors.initials: A
authors.lastname: Haghparast
mesh_terms: D052157:Aptamers, Nucleotide / Q000096:biosynthesis* / Q000737:chemistry; D001943:Breast Neoplasms / Q000175:diagnosis*; D045744:Cell Line, Tumor; D015342:DNA Probes / Q000096:biosynthesis* / Q000737:chemistry; D005260:Female; D006801:Humans; D008024:Ligands; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000032:analysis* / Q000737:chemistry / Q000235:genetics; D052156:SELEX Aptamer Technique
chemical_list: D052157:Aptamers, Nucleotide; D015342:DNA Probes; D008024:Ligands; D018719:Erb-b2 Receptor Tyrosine Kinases; C508053:ERBB2 protein, human