期刊
BIOSENSORS & BIOELECTRONICS
卷 41, 期 -, 页码 477-483出版社
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2012.09.012
关键词
Macroporous silicon; Fluorescence quenching; Simple one-step assay
类别
资金
- NLRL (National Leading Research Lab.) [2012R1A2A1A01008085]
- Public welfare & Safety research program [2010-0020778]
- Basic Science Research Program (ERC program) of the National Research Foundation of Korea (NRF) [2010-0029409]
- NRF [2010-0027771, R0A-2008-000-20078-0]
- Korea government (MEST)
- Seoul RBD Program [PA100026M0211612]
- National Research Foundation of Korea [R0A-2008-000-20078-0] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
We synthesized 3D macroporous silicon through a simple electrochemical dissolution process and systematically estimated its protein adsorption and effect on fluorescence emission. Compared with conventional 2D polystyrene plate, the macroporous silicon showed a superior protein adsorption capacity and significant fluorescence quenching effect. We developed a 3D macroporous silicon-based adenosine assay system through the following fabrication process: streptavidin molecules that have been immobilized on the surface of macroporous silicon are attached with biotin-linked and adenosine-specific DNA aptamer, followed by hybridization between the attached aptamer and fluorescent chemical (carboxytetramethylrhodamine/CTMR) that is conjugated with a short complementary DNA sequence. In the absence of adenosine, the aptamer-CTMR complexes remain closely attached to the surface of porous silicon, hence fluorescence being significantly quenched. Upon binding to adenosine, the DNA aptamer is subject to structure switching that leads to dissociation of CTMR from DNA aptamer, and consequently the CTMR fluorescence is restored, indicating a simple one-step assay of adenosine. Compared to the conventional 2D PS and ZnO nanorods-based assays, adenosine at much lower (sub-micromolar) concentration was successfully detected through the 3D macroporous silicon-based assay. The three-dimensionally and densely immobilized aptamer probes and effective fluorescence quenching on the surface of macroporous silicon enables adenosine to be detected at lower levels. Although the adenosine detection is reported here as a proof-of-concept, the developed macroporous silicon-based simple one-step assay platform can be applied in general to fluorescence quenching -based detection of many other biomolecules. (C) 2012 Elsevier B.V. All rights reserved.
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