Antigen peptide-based immunosensors for rapid detection of antibodies and antigens.
Antigen peptide-based immunosensors for rapid detection of antibodies and antigens.
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DOI：
10.1021/ac900845a
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发表时间：
2009-07-01
影响因子：
7.4
通讯作者：
Heyduk, Tomasz
中科院分区：
文献类型：
--
作者：
Tian, Ling;Heyduk, Tomasz
这里描述的均质免疫传感器设计利用抗体的二价性质。抗原肽使用柔性接头与短互补寡核苷酸（信号寡核苷酸）缀合，所述短互补寡核苷酸各自含有可形成荧光共振能量转移（FRET）供体-受体对的荧光染料。互补信号寡核苷酸足够短以防止它们自身退火。肽-信号寡核苷酸构建体与二价抗体的结合导致信号寡核苷酸的局部浓度的大幅增加，引起它们的退火和FRET信号的出现。我们使用简单的模型系统（抗生物素抗体）来获得传感器设计的原理验证。然后，我们构建了两个传感器的基础上的两个肽对应的抗原的两个抗体提出的人心肌肌钙蛋白I。我们证明了这些传感器可用于抗体的灵敏检测和完整肌钙蛋白I的竞争检测。此外，我们表明，这些传感器可用于检测激酶活性靶向抗原肽。这些简单而强大的免疫传感器可以应用于抗体检测（例如，在自身免疫或传染病的诊断），蛋白质检测（特别是当检测速度是必不可少的），并在检测蛋白质的翻译后修饰所涉及的酶活性的测定。
The homogenous immunosensor design described here utilizes bivalent nature of the antibody. Antigen peptide is conjugated using flexible linkers with short complementary oligonucleotides (signaling oligonucleotides) containing each a fluorochrome that can form a Fluorescence Resonance Energy Transfer (FRET) donor-acceptor pair. The complementary signaling oligonucleotides are short enough to prevent their annealing on their own. Binding of the peptide-signaling oligonucleotide constructs to bivalent antibody results in a large increase in local concentration of signaling oligonucleotides causing their annealing and appearance of FRET signal. We used simple model system (anti-biotin antibody) to obtain proof-of-principle validation of the sensor design. We then constructed two sensors based on two peptides corresponding to the antigens of two antibodies raised against human cardiac troponin I. We demonstrated that these sensors could be used for sensitive detection of the antibody and for competition-based detection of the intact troponin I. Furthermore, we showed that these sensors could be used for detection of kinase activity targeting the antigen peptide. These simple and robust immunosensors may find applications in antibody detection (for example, in diagnosis of autoimmune or infectious disease), in protein detection (especially when speed of detection is essential), and in assays for detecting enzymatic activities involved in posttranslational modifications of proteins.
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影响因子：
4.8
作者：
Heyduk, E;Heyduk, T;Wisniewski, JR
通讯作者：
Wisniewski, JR
影响因子：
3.5
作者：
MITTMANN, K;JAQUET, K;HEILMEYER, LMG
通讯作者：
HEILMEYER, LMG
影响因子：
3.9
作者：
Perry, S. V.
通讯作者：
Perry, S. V.
影响因子：
4.8
作者：
Ferrer-Miralles, N;Feliu, JX;Villaverde, A
通讯作者：
Villaverde, A
DOI：
10.1007/10_2007_075
发表时间：
2008-01-01
期刊：
BIOSENSING FOR THE 21ST CENTURY
影响因子：
--
作者：
Chan, Cangel Pui-yee;Cheung, Yiu-chi;Seydack, Matthias
通讯作者：
Seydack, Matthias