Mulitplexed protein measurement by real-time immuno-PCR

描述（由申请人提供）：实时聚合酶链反应（PCR）是定量基因表达谱最常用的方法；它可以同时对数百个mRNA转录物进行极其灵敏的测量。我们提出了一种方法，通过该方法，蛋白质丰度可以定量使用相同的技术。这是通过实施三明治免疫分析格式实现的，其中检测抗体被标记为不同的DNA序列。实时聚合酶链式反应（Real-time PCR）用于同时定量所有DNA标记，从而揭示生物样品中同源蛋白的数量。初步实验表明，与标准ELISA相比，使用这种抗体- dna偶联物的实时PCR检测灵敏度提高了两个数量级以上，动态范围增加了近三个数量级，并且标准偏差非常低（CV约为5%）。也获得了支持该方法复用能力的初步证据。

DESCRIPTION (provided by applicant): The real-time polymerase chain reaction (PCR) is the most commonly used method for quantitative gene expression profiling; it allows extremely sensitive measurement of hundreds of mRNA transcripts in parallel. We propose a method by which protein abundances can be quantitated using the same technology. This is achieved by implementing a sandwich immuno-assay format in which the detection antibodies are labeled with different DNA sequences. Real-time PCR is then used to quantify all of the DNA labels in parallel, thus revealing the quantities of the cognate proteins in the biological samples. Preliminary experiments show that real-time PCR using such antibody-DNA conjugates gives over two orders of magnitude more sensitive detection and almost three orders of magnitude increased dynamic range in comparison to standard ELISA, and standard deviations are very low (CV approximately 5%). Initial evidence supporting the ability to multiplex this method has also been obtained.