期刊
BIOSENSORS & BIOELECTRONICS
卷 26, 期 7, 页码 3233-3239出版社
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2010.12.032
关键词
Silicon nanowire; Biosensor; Self-assembled monolayer; Interactions; Protein; DNA
类别
资金
- Agency for Science, Technology and Research (A*STAR), Singapore [CCOG01-005-2008]
The large number of estrogen receptor (ER) binding sites of various sequence patterns requires a sensitive detection to differentiate between subtle differences in ER-DNA binding affinities. A self-assembled monolayer (SAM)-assisted silicon nanowire (SiNW) biosensor for specific and highly sensitive detection of protein-DNA interactions, remarkably in nuclear extracts prepared from breast cancer cells, is presented. As a typical model, estrogen receptor element (ERE, dsDNA) and estrogen receptor alpha (ER alpha, protein) binding was adopted in the work. The SiNW surface was coated with a vinyl-terminated SAM, and the termination of the surface was changed to carboxylic acid via oxidation. DNA modified with amine group was subsequently immobilized on the SiNW surface. Protein-DNA binding was finally investigated by the functionalized SiNW biosensor. X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM) were employed to characterize the stepwise functionalization of the SAM and DNA on bare silicon surface, and to visualize protein-DNA binding on the SiNW surface, respectively. We observed that ER alpha had high sequence specificity to the SiNW biosensor which was functionalized with three different EREs including wild-type, mutant and scrambled DNA sequences. We also demonstrate that the specific DNA-functionalized SiNW biosensor was capable of detecting ER alpha as low as 10 fM. Impressively, the developed SiNW biosensor was able to detect ER alpha-DNA interactions in nuclear extracts from breast cancer cells. The SAM-assisted SiNW biosensor, as a label-free and highly sensitive tool, shows a potential in studying protein-DNA interactions. (C) 2010 Elsevier B.V. All rights reserved.
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