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RNA and DNA aptamers in cytomics analysis
Author(s) -
Ulrich Henning,
Martins Antonio Henrique B.,
Pesquero João Bosco
Publication year - 2004
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.20056
Subject(s) - aptamer , systematic evolution of ligands by exponential enrichment , microbiology and biotechnology , biology , monoclonal antibody , rna , computational biology , dna , flow cytometry , western blot , antibody , biochemistry , gene , genetics
Background The systematic evolution of ligands by exponential enrichment (SELEX) technique is a combinatorial library approach in which DNA or RNA molecules (aptamers) are selected by their ability to bind their protein targets with high affinity and specificity, comparable to that of monoclonal antibodies. In contrast to antibodies conventionally selected in animals, aptamers are generated by an in vitro selection process, and can be directed against almost every target, including antigens like toxins or nonimmunogenic targets, against which conventional antibodies cannot be raised. Methods Aptamers are ideal candidates for cytomics, as they can be attached to fluorescent reporters or nanoparticles in order to study biological function by fluorescence microscopy, by flow cytometry, or to quantify the concentration of their target in biological fluids or cells using ELISA, RIA, and Western blot assays. Results We demonstrate the in vitro selection of anti‐kinin B1 receptor aptamers that could be used to determine B1 receptor expression during inflammation processes. These aptamers specifically recognize their target in a Northern‐Western blot assay, and bind to their target protein whenever they are exposed in the membrane. Conclusions Currently, aptamers are linked to fluorescent reporters. We discuss here the present status and future directions concerning the use of the SELEX technique in cytomics. © 2004 Wiley‐Liss, Inc.