Integrated poly(dimethysiloxane) with an intrinsic nonfouling property approaching "absolute" zero background in immunoassays.
Integrated poly(dimethysiloxane) with an intrinsic nonfouling property approaching "absolute" zero background in immunoassays.
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DOI：
10.1021/ac101277e
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发表时间：
2010-08
影响因子：
7.4
通讯作者：
Hongwei Ma;Yuanzi Wu;Xiaoli Yang;Xing Liu;Jian’an He;Long Fu;Jie Wang;Hongke Xu;Y. Shi;R. Zhong
中科院分区：
文献类型：
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作者：
Hongwei Ma;Yuanzi Wu;Xiaoli Yang;Xing Liu;Jian’an He;Long Fu;Jie Wang;Hongke Xu;Y. Shi;R. Zhong
实现高灵敏度和特异性的蛋白质微阵列分析的关键是防止非特异性蛋白质吸附到“绝对”零水平，因为任何信号放大方法都会同时放大信号和噪声。在这里，我们开发了一种新的固体支撑材料，即聚合物涂覆的引发剂集成的聚（二甲基硅氧烷）（iPDMS），其能够实现这种“绝对”零（即，低于仪器的检测限）。该iPDMS的实施使得能够对11种肿瘤标志物进行实用且高质量的多重酶联免疫吸附测定（ELISA）。该iPDMS不需要任何封闭步骤，仅需要温和的洗涤条件。与主流的硝酸纤维素（NC）薄膜相比，它使用的捕获抗体平均少8倍。除了节省时间和材料外，iPDMS的检测限（LOD）低至19 pg/mL（-1），这对于目前大多数临床诊断应用来说是足够低的。我们期望看到这种iPDMS对实现蛋白质微阵列在研究和实际应用中的巨大潜力的直接影响，例如大规模和高通量筛选，临床诊断，检验和检疫。
The key to achieve a highly sensitive and specific protein microarray assay is to prevent nonspecific protein adsorption to an "absolute" zero level because any signal amplification method will simultaneously amplify signal and noise. Here, we develop a novel solid supporting material, namely, polymer coated initiator integrated poly(dimethysiloxane) (iPDMS), which was able to achieve such "absolute" zero (i.e., below the detection limit of instrument). The implementation of this iPDMS enables practical and high-quality multiplexed enzyme-linked immunosorbent assay (ELISA) of 11 tumor markers. This iPDMS does not need any blocking steps and only require mild washing conditions. It also uses on an average 8-fold less capture antibodies compared with the mainstream nitrocellulose (NC) film. Besides saving time and materials, iPDMS achieved a limit-of-detection (LOD) as low as 19 pg mL(-1), which is sufficiently low for most current clinical diagnostic applications. We expect to see an immediate impact of this iPDMS on the realization of the great potential of protein microarray in research and practical uses such as large scale and high-throughput screening, clinical diagnosis, inspection, and quarantine.