Journal
BIOSENSORS & BIOELECTRONICS
Volume 39, Issue 1, Pages 44-50Publisher
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2012.06.038
Keywords
Label-free sensor; Aptasensor; Aptamer; Aptaswitch; Modular aptamer
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Funding
- Converging Research Center Program through the National Research Foundation of Korea (NRF)
- Ministry of Education, Science and Technology [2011K000841]
- Korea Biotech R&D Group of Next-generation Growth Engine Project of the Ministry of Education, Science and Technology, Republic of Korea [M10417060004-04N1706-00410]
- National Research Foundation of Korea [2010-50206] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
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An aptamer can be redesigned to new functional molecules by conjugating with other oligonucleotides. However, it requires experimental trials to optimize the conjugating module with the sensitivity and selectivity toward a target. To reduce these efforts, we report rationally-designed modular allosteric aptamer sensor (MAAS), which is composed of coupled two aptamers and the regulator. For label-free protein detection, the protein-aptamer was conjugated with the malachite green (MG) aptamer for signaling. The MAAS additionally has the regulator domain which is designed to hybridize to a protein binding domain. The regulator makes MAAS to be inactive by destructing the original structure of the two aptamers. However, its conformation becomes active by dissociating the hybridization from the protein recognition signal, thereby inducing the binding of MG emitting the enhanced fluorescence. The design of regulator is based on the thermodynamic energy difference by the RNA conformational change and protein-aptamer affinity. Here we first demonstrated the MAAS for hepatitis C helicase and replicase. The target proteins were detected up to 250 nM with minimized blank signals and displayed high specificities 10-fold greater than in non-specific proteins. The MAAS provides valuable tools that can be adapted to a wide range of configurations in bioanalytical applications. (c) 2012 Elsevier B.V. All rights reserved.
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