Peptide detection using the proximity ligation assay

描述(由申请人提供)： 蛋白质组分析正在给人类疾病的诊断带来革命性的变化。现有技术能够精确定位指示疾病进展的蛋白质，使其能够用于早期检测和治疗方案。然而，蛋白质组技术的使用受到限制，因为它们的使用和数据解释的复杂性。作为当前多肽和蛋白质分析方法的替代方法，我建议在邻近连接分析中使用新型多肽特异性核酸适体来检测复杂混合物中的独特多肽。在这个系统中，一对适配子将识别多肽的末端，并凭借它们的邻近性被酶连接在一起。因为大多数多肽是由它们的末端序列唯一指定的，所以连接产物指示一个独特的多肽。多肽的存在可以通过聚合酶链式反应来评估，而不是通过质量鉴定。使用这种新的方法，人们可以量化多肽浓度，识别低丰度多肽，并可能在单一反应中检测翻译后修饰。我们设想将该方法应用于通过微阵列读出，将复杂的蛋白质混合物的测量简化到单芯片平台。

DESCRIPTION (provided by applicant): Proteomic analyses are revolutionizing the diagnosis of human disease. Existing technologies are able to pinpoint the proteins indicative of disease progression, enabling their use in early detection and treatment scenarios. However, the use of proteomic techniques is limited because of the complexities of their usage and data interpretation. As an alternative to the current methods of peptide and protein analysis, I propose employing novel peptide-specific nucleic acid aptamers in the Proximity Ligation Assay for the detection of unique peptides in complex mixtures. In this system, a pair of aptamers would recognize the terminal ends of a peptide and, by virtue of their proximity, be enzymatically ligated together. Because most peptides are uniquely specified by their terminal sequences, the ligation product indicates a unique peptide. The presence of a peptide can be assessed via a PCR reaction, instead of its identification by mass. Using this novel method, one could quantitate peptide concentrations, identify low-abundance peptides and detect post-translation modifications, potentially in a single reaction. We envision adapting the method to readout via a microarray, simplifying complex protein mixture measurements to a single chip platform.