Generation of an Enriched Pool of DNA Aptamers for a HER2 Overexpressing Cell Line selected by Cell –SELEX

نویسنده:
, , , , , , ,
سال
: 2011
چکیده: 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.
کلیدواژه(گان): Keywords: aptamer,breast cancer,Cell ELA,Cell SELEX,HER2
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