Polymer-based elemental tags for sensitive Bioassays
Polymer-based elemental tags for sensitive Bioassays
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DOI：
10.1002/anie.200700796
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发表时间：
2007-01-01
影响因子：
16.6
通讯作者：
Winnik, Mitchell A.
中科院分区：
文献类型：
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作者：
Lou, Xudong;Zhang, Guohua;Winnik, Mitchell A.
为了识别复杂混合物中的稀有（例如患病或外来）细胞，或了解细胞的蛋白质组复杂性 [1, 2]，需要能够同时定量地测量复杂样品中可能存在的大量蛋白质或其他生物标志物。这是一项艰巨的任务，超出了目前的能力范围。为了解决这种复杂性的问题，我们已经开始开发一种基于元素标签的高灵敏度测定方法 [3-6]，该方法可以同时测量单个样品中的多种蛋白质。这种方法的优点在于生物系统中存在大量低丰度的可用元素和同位素（可能超过 79 种），这将允许同时使用多个标签。电感耦合等离子体质谱 (ICP-MS) 是检测和量化这些标签的理想技术，因为 ICP-MS 提供了标签质量之间出色的分辨率和出色的动态范围（九个数量级）。 [7]这种方法可以克服当前可用的荧光标记方法的一些局限性。 [8]这些限制源于不同染料的光谱重叠以及同时测量丰度相差一个数量级或更多的目标的困难。 ICP-MS 检测的其他优点包括灵敏度高，可与放射免疫分析或化学发光分析相媲美，[3] 元素标签对光漂白和存储时间不敏感，以及标记样品的稳定性，以便可以存储或运输用于分析。我们在此讨论用于 ICP-MS 检测的新型元素标签的开发以及它们在标记抗体中的用途，这些抗体被选择以允许特异性识别独特的细胞表面标记。通过使用这种技术，应该可以达到十亿分之一的检测限，这将允许检测拷贝数低至 100 的细胞表面标记。
To identify a rare (eg, diseased or foreign) cell in a complex mixture, or to understand the proteomic complexity [1, 2] of cells, one needs to be able to measure simultaneously and quantitatively a large number of proteins or other biomarkers that may be present in a complex sample. This is a difficult task and is beyond the reach of current capabilities. To address a problem of this complexity, we have begun to develop a high-sensitivity assay [3–6] based upon elemental tags that will enable the simultaneous measurement of many proteins in a single sample. The advantage of this approach lies in the large number of available elements and isotopes (potentially greater than 79) found in low abundance in biological systems, which will allow multiple tags to be used simultaneously. Inductively coupled plasma mass spectrometry (ICP-MS) is an ideal technique for detecting and quantifying these tags, as ICP-MS provides excellent resolution between the tag masses and an exceptional dynamic range (nine orders of magnitude).[7] This method allows one to overcome some of the limitations of currently available fluorescent tagging approaches.[8] These limitations arise from the spectral overlap of different dyes and the difficulty in measuring simultaneously targets that differ in abundance by an order of magnitude or more. Other benefits of ICP-MS detection include the high sensitivity, which is comparable to that of radioimmunoassays or chemiluminescent assays,[3] insensitivity of elemental tags to photobleaching and storage time, as well as the stability of the tagged sample so that it can be stored or shipped for analysis. We discuss herein the development of a new class of elemental tags for ICP-MS detection and their use for tagging of antibodies chosen to allow specific recognition of distinguishing cell surface markers. By using this technique it should be possible to achieve detection limits on the order of parts per billion, which will allow the detection of cell surface markers with copy numbers as low as 100.