Human Proteome Arrays for Auto-Antibody Identification in Clinical Cancer Studies

描述（由申请人提供）：用于临床癌症研究中自身抗体鉴定的人类蛋白质组阵列本项目的目标是在蛋白质组规模上开发蛋白质阵列，用于癌症特异性自身抗体特征的高通量发现和验证。早期发现癌症是成功治疗癌症患者的关键因素。癌症患者中特异性自身抗体特征的发现导致了对自身抗体应答可能具有诊断潜力的认识。用于快速筛选和验证多种感兴趣抗原的高通量方法的缺乏已被确定为开发自身抗体作为癌症诊断的生物标志物的重要瓶颈。我们将测试正常和癌症患者的血清对蛋白质阵列产生的基因文库表达人类全长蛋白质。针对这些阵列中蛋白质的自身抗体将使用电化学发光（ECL）检测技术进行鉴定，这是一种经过验证的高通量基于阵列的测量技术，可以对大量患者样本进行研究。我们的方法将提供一个高通量，灵敏和特异性的检测平台，提供现有的基于阵列的方法的关键优势。这种方法还将促进从使用大阵列发现抗原到在同一诊断平台上使用小聚焦阵列对命中进行临床验证的快速转变。在拟议项目的第一阶段，我们将证明该方法的技术可行性;在第二阶段，我们将开发和验证蛋白质组规模的阵列，并使用它们来筛选癌症血清样本，以识别有希望的自身抗原子集;然后将用大量的个体样本进一步测试选定的子集。

DESCRIPTION (provided by applicant): Human Proteome Arrays for Auto-Antibody Identification in Clinical Cancer Studies The goal of this project is to develop protein arrays on a proteome scale for high throughput discovery and validation of cancer-specific autoantibody signatures. Early detection of cancer is a critical factor for the successful treatment of cancer patients. The discovery of specific autoantibody signatures in cancer patients has led to the recognition that autoantibody responses may have diagnostic potential. The lack of high throughput methods for rapidly screening and validating multiple antigens of interest has been identified as an important bottleneck for the development of autoantibodies as biomarkers for cancer diagnosis. We will test normal and cancer patient sera against protein arrays generated using a gene library expressing human full-length proteins. Autoantibodies against the proteins in these arrays will be identified using electrochemiluminescence (ECL) detection technology, a proven technology for high throughput array-based measurements that will enable studies with large numbers of patient samples. Our approach will offer key advantages over existing array-based approaches by providing a high throughput, sensitive and specific assay platform. This approach will also facilitate the rapid transition from discovery of antigens using large arrays to clinical validation of the hits using small focused arrays on the same diagnostic platform. In the Phase I portion of the proposed project, we will demonstrate technical feasibility of the approach; in the Phase II portion we will develop and validate the proteome-scale arrays, and use them to screen cancer serum samples to identify a promising subset of autoantigens; the selected subset will then be further tested with a large number of individual samples.