Two-Dimensional Electrophoresis-Based Selection of Aptamers Against an Unidentified Protein in a Tissue Sample

Autor:	Shinichi Goto, Nasa Savory, Kazunori Ikebukuro, Koichi Abe, Wataru Yoshida, Yutaka Unuma, Stefano Ferri, Makoto Nakamura
Rok vydání:	2013
Předmět:	Chemistry Oligonucleotide Aptamer Biochemistry (medical) Clinical Biochemistry Proteomics Biochemistry Molecular biology Analytical Chemistry Blot chemistry.chemical_compound Electrochemistry Target protein Biomarker discovery Spectroscopy DNA Systematic evolution of ligands by exponential enrichment
Zdroj:	Analytical Letters. 46:2954-2963
ISSN:	1532-236X 0003-2719
DOI:	10.1080/00032719.2013.828300
Popis:	A fully automated two-dimensional electrophoresis (2DE) system was employed for DNA aptamer selection against an unidentified protein in a mouse liver tissue extract as a model target. A 2DE-based systematic evolution of ligands by exponential enrichment (2DE-SELEX) was demonstrated for aptamer selection against a single protein spot that was separated on a nitrocellulose membrane. After four iterative 2DE-SELEX cycles, the oligonucleotide pool was sequenced and aptamer sequences were identified. A blotting assay showed that an identified aptamer with a stable stem–loop structure had specific binding activity against the target protein. The 2DE-SELEX was shown to be promising for the development of aptamers against unidentified proteins in complex samples for proteomic analysis and biomarker discovery. Supplemental materials are available for this article. Go to the publisher's online edition of Analytical Letters to view the supplemental file.
Databáze:	OpenAIRE
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