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In Vitro Selection of Aptamers with Affinity for Neuropeptide Y Using Capillary Electrophoresis

2005; American Chemical Society; Volume: 127; Issue: 26 Linguagem: Inglês

10.1021/ja052406n

1943-2984

Shaun D. Mendonsa, Michael T. Bowser,

Biosensors and Analytical Detection

Capillary electrophoresis-systematic evolution of ligands by exponential enrichment (CE-SELEX) was used to select aptamers for neuropeptide Y (NPY). This is the first example of a CE-SELEX selection for aptamers that bind a target molecule smaller than itself. One of the limitations of CE-SELEX is that the aptamer must exhibit a significant mobility shift when it binds the target to facilitate fraction collection. Before this study, it was not clear if smaller targets would be capable of inducing a large enough shift in mobility for CE-SELEX to be successful. NPY is a 36-amino acid peptide (MW = 4272 g/mol), much smaller than the 80-base ssDNA used in the selection (∼25 kDa). NPY binding aptamers with 300−1000 nM dissociation constants were obtained after only four rounds of selection. The specificity of the aptamers was tested using human pancreatic polypeptide (hPP). hPP is a 36-amino acid peptide with ∼50% homology with NPY. Aptamers with up to 42-fold selectivity for NPY over hPP were observed.