期刊
BIOSENSORS & BIOELECTRONICS
卷 51, 期 -, 页码 255-260出版社
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2013.08.001
关键词
Proximity effect; Enzyme-assisted signal amplification; Protein detection
类别
资金
- National Natural Science Foundation of China [20905023, 21175035, 21190044]
- National Basic Research Program [2011CB911002]
- International Science & Technology Cooperation Program of China [2010DFB30300]
- Hunan Province Science and Technology Project of China [2010SK3002, 2013FJ4042]
In this paper, we develop a sensitive fluorescence method for protein detection based on proximity extension and enzyme-assisted signal amplification. In this novel method, pairs of proximity probes are designed, and the recognition elements are integrated into the proximity probes. Then proteins are detected by transforming aptamer or antibody-protein binding signals into DNA detection based on proximity effect. In addition, nick sites are introduced into the proximity probes to amplify the detectable signal. As proof of concept, detection of human a-thrombin and human IgG are demonstrated in this study. The aptamers and antibodies are coupled in the proximity probes as recognition elements for human a-thrombin and human IgG respectively. In the presence of target protein, aptamer or antibody-protein binding signals are transformed into detectable signals by the proximity effect, and can be further amplified by enzyme-assisted strand displacement. The above mentioned strategies consequently bring the limit of detection (LOD) to as low as 1 pM for human a-thrombin and 6 pM for human IgG. Furthermore, this method might be extended to sensitive detection of other proteins by changing recognition elements of proximity probes. (C) 2013 Elsevier B.V. All rights reserved.
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