Infrared-based protein detection arrays for quantitative proteomics
Infrared-based protein detection arrays for quantitative proteomics
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DOI：
10.1002/pmic.200600757
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发表时间：
2007-02-01
期刊：
影响因子：
3.4
通讯作者：
Korf, Ulrike
中科院分区：
文献类型：
--
作者：
Loebke, Christian;Sueltmann, Holger;Korf, Ulrike
为了满足系统生物学和药物发现的需要而发展有效的技术，到目前为止还没有得到足够的重视。对信号网络进行时间分辨定量描述的一个实质性瓶颈是目前建立的方法的有限吞吐量和灵敏度不足。在这里，我们提出了一种改进的基于蛋白质微阵列的方法，该方法基于近红外范围内的信号检测，可以对fg范围内的蛋白质进行敏感检测。该试验的高灵敏度允许仅从20,000个细胞中提取蛋白质的特定定量，错误率仅为5%。容量仅限于分析多达500个不同的样品每个微阵列。蛋白质丰度是定性和定量确定，如果重组蛋白是可用的。这种新方法被称为IPAQ（基于红外的定量读出蛋白阵列）。IPAQ提供了一种高灵敏度的实验方法，优于现有的标准蛋白质定量技术，适用于定量蛋白质组学。采用IPAQ方法，可以很容易地对活检样本中激活的信号网络和药物发现策略所需的信号模块之间的串扰进行详细分析。
The advancement of efficient technologies to comply with the needs of systems biology and drug discovery has so far not received adequate attention. A substantial bottleneck for the time-resolved quantitative description of signaling networks is the limited throughput and the inadequate sensitivity of currently established methods. Here, we present an improved protein microarray-based approach towards the sensitive detection of proteins in the fg-range which is based on signal detection in the near-infrared range. The high sensitivity of the assay permits the specific quantification of proteins derived from as little as only 20 000 cells with an error rate of only 5%. The capacity is limited to the analysis of up to 500 different samples per microarray. Protein abundance is determined qualitatively, and quantitatively, if recombinant protein is available. This novel approach was called IPAQ (infrared-based protein arrays with quantitative readout). IPAQ offers a highly sensitive experimental approach superior to the established standard protein quantification technologies, and is suitable for quantitative proteomics. Employing the IPAQ approach, a detailed analysis of activated signaling networks in biopsy samples and of crosstalk between signaling modules as required in drug discovery strategies can easily be performed.