Protein detection using biobarcodes.
Protein detection using biobarcodes.
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DOI：
10.1039/b608442g
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发表时间：
2006-09
影响因子：
--
通讯作者：
U. Müller
中科院分区：
文献类型：
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作者：
U. Müller
在过去的50年里，检测蛋白质分析物的检测方法的发展一直受到对更高水平灵敏度和多路复用的持续需求的推动。结果是三明治型免疫测定的发展，从简单的放射性同位素，比色或荧光标记系统开始，包括各种酶或纳米结构的信号放大方案，伴随灵敏度增加超过100万倍。样品和测试的多路复用已经分别通过微孔板和微阵列平台实现，或者最近通过各种分子条形码系统实现。通过将核酸扩增步骤与标准的双边免疫测定相结合，两种不同的平台已成为目前的领跑者。在这两种方法中，捕获的蛋白质分析物被作为替代靶标的多种寡核苷酸所取代。其中一个平台使用DNA或RNA聚合酶进行扩增步骤，而检测是通过荧光。另一种是基于金纳米粒子的扩增和检测。后一种技术，现在被称为生物条形码，完全不含酶，并提供潜在的更高的多路复用能力。
Over the past 50 years the development of assays for the detection of protein analytes has been driven by continuing demands for higher levels of sensitivity and multiplexing. The result has been a progression of sandwich-type immunoassays, starting with simple radioisotopic, colorimetric, or fluorescent labeling systems to include various enzymatic or nanostructure-based signal amplification schemes, with a concomitant sensitivity increase of over 1 million fold. Multiplexing of samples and tests has been enabled by microplate and microarray platforms, respectively, or lately by various molecular barcoding systems. Two different platforms have emerged as the current front-runners by combining a nucleic acid amplification step with the standard two-sided immunoassay. In both, the captured protein analyte is replaced by a multiplicity of oligonucleotides that serve as surrogate targets. One of these platforms employs DNA or RNA polymerases for the amplification step, while detection is by fluorescence. The other is based on gold nanoparticles for both amplification as well as detection. The latter technology, now termed Biobarcode, is completely enzyme-free and offers potentially much higher multiplexing power.