Document Detail

Nanoparticle-aptamer conjugates for cancer cell targeting and detection.
MedLine Citation:
PMID:  20217600     Owner:  NLM     Status:  MEDLINE    
Aptamers are DNA or RNA oligonucleotide sequences that selectively bind to their target with high affinity and specificity. They are obtained using an iterative selection protocol called SELEX. Several small molecules and proteins have been used as targets. Recently, a variant of this methodology, known as cell-SELEX, has been developed for a new generation of aptamers, which are capable of recognizing whole living cells. We have used this methodology for the selection of aptamers, which show high affinity and specificity for several cancer cells. In this chapter, we describe (1) the process followed for the generation of aptamers capable of recognizing acute leukemia cells (CCRF-CEM cells) and (2) the method of enhancing the selectivity and sensitivity of these aptamers by conjugation with a dual-nanoparticle system, which combines magnetic nanoparticles (MNP) and fluorescent silica nanoparticles (FNP). Specifically, the selected aptamers, which showed dissociation constants in the nanomolar range, have been coupled to MNPs in order to selectively collect and enrich cells from complex matrices, including blood samples. The additional coupling of the aptamer to FNPs offers an excellent and highly sensitive method for detecting cancer cells. In order to prove the potential of this rapid and low-cost method for diagnostic purposes, confocal microscopy was used to confirm the specific collection and detection of target cells in concentrations as low as 250 cells. The final fluorescence of the cells labeled with the nanoparticles was quantified using a fluorescence microplate reader.
M Carmen Est?vez; Yu-Fen Huang; Huaizhi Kang; Meghan B O'Donoghue; Suwussa Bamrungsap; Jilin Yan; Xiaolan Chen; Weihong Tan
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  624     ISSN:  1940-6029     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2010  
Date Detail:
Created Date:  2010-03-10     Completed Date:  2010-06-14     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  235-48     Citation Subset:  IM    
Department of Chemistry, Functional Genomics Shands Cancer Center, Center for Research at the Bio/Nano Interface, University of Florida, Gainesville, FL, USA.
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MeSH Terms
Aptamers, Nucleotide / diagnostic use*
Cell Line, Tumor
Limit of Detection
Microscopy, Confocal
Nanomedicine / methods*
Nanoparticles / diagnostic use*
Neoplasms / diagnosis*,  pathology*
SELEX Aptamer Technique / methods*
Reg. No./Substance:
0/Aptamers, Nucleotide

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