Selection of aptamers for molecular recognition and characterization of cancer cells.
In this paper, we describe a new way to generate molecular probes for specific recognition of cancer cells. Molecular medicine will require a large number of probes for molecular recognition and characterization of a variety of diseased cells. Aptamers, single-stranded DNA/RNA probes, are poised to become a chemist's antibody and have the potential to serve as molecular probes for a variety of biomedical applications. By applying newly developed cell-SELEX (cell-based systematic evolution of ligands by exponential enrichment) against whole living cells, panels of aptamers have been evolved from an initial DNA library to characterize target cells at the molecular level. Ramos cells, a B-cell lymphoma cell line, were used as target cells for the generation of effective molecular probes. By taking advantages of the repetitive and broad enrichment strategy, the selected aptamers could bind to target cells and other closely related cell lines in variant patterns with an equilibrium dissociation constant (Kd) in the nanomolar range. Some aptamers could also specifically recognize the target lymphoma cells mixed with normal human bone marrow aspirates. The cell-based SELEX is simple, fast, and robust. The strategies used here will be highly useful for aptamer selection against complex target samples in order to generate a large number of aptamers in a variety of biomedical and biotechnological applications, paving the way for molecular diagnosis, therapy, and biomarker discovery.
Tang
Zhiwen
Z
Department of Chemistry, Shands Cancer Center, Center for Research at Bio/nano Interface, UF Genetics Institute, University of Florida, Gainesville, Florida 32611-7200, USA.
Shangguan
Dihua
D
Wang
Kemin
K
Shi
Hui
H
Sefah
Kwame
K
Mallikratchy
Prabodhika
P
Chen
Hui William
HW
Li
Ying
Y
Tan
Weihong
W
D052157:Aptamers, Nucleotide / Q000032:analysis*; D001483:Base Sequence; D001665:Binding Sites; D014408:Biomarkers, Tumor / Q000032:analysis*; D001854:Bone Marrow Cells / Q000737:chemistry* / Q000473:pathology; D045744:Cell Line, Tumor; D015342:DNA Probes / Q000032:analysis; D019286:Endopeptidase K / Q000378:metabolism; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate; D006801:Humans; D008223:Lymphoma / Q000737:chemistry / Q000473:pathology*; D016393:Lymphoma, B-Cell / Q000737:chemistry / Q000473:pathology; D015347:RNA Probes / Q000032:analysis; D052156:SELEX Aptamer Technique / Q000379:methods*; D014357:Trypsin / Q000378:metabolism
D052157:Aptamers, Nucleotide; D014408:Biomarkers, Tumor; D015342:DNA Probes; D015347:RNA Probes; D014357:Trypsin; D019286:Endopeptidase K; D016650:Fluorescein-5-isothiocyanate