APT: the Analytical Proteomics Team

描述（由申请人提供）：生物标志物鉴定和表征对于更精确的诊断和定制治疗具有很大的希望。人类癌症的异质性和这些疾病中未满足的医疗需求为关注癌症生物标志物的开发提供了令人信服的论据。基于质谱（MS）的蛋白质组学方法以飞摩尔灵敏度和高分析精度提供了对癌症和其他疾病的生物标志物的深入了解，但由于人类生物流体的复杂性和这些方法中采用的方案，结果已被证明难以再现。该CPTAC提案将为采用电喷雾电离（ESI）和基质辅助激光解吸电离（MALDI）平台的MS蛋白质组学开发强大的协议和标准。离子迁移率MS提供了几个数量级的增加动态范围，并将进一步探索与这个新兴的技术平台的癌症生物标志物候选人的检测协议。该联盟将采用高特异性免疫试剂开发平台，用于精确检测和定量乳腺癌和前列腺癌相关生物标志物。特定生物标志物蛋白的免疫亲和选择将在特定蛋白的MS检测和定量之前使用纳米级免疫亲和柱作为样品分级分离步骤来实现。一种新兴的技术，将特异性抗体结合在微制造的“生物CD”上，通过旋转圆盘干涉法读取，能够在几分钟内对数百个样品中的数百种分析物进行无标记评估。我们将集中在候选生物标志物相关的乳腺癌和前列腺癌从NF？B和STATS信号通路。三个营利性和一个非营利企业合作伙伴将加入几个在蛋白质组学技术方面拥有深厚专业知识和广泛资源的学术团体，以最有效地评估和推出MS和亲和蛋白质组学方法的强大协议和标准。前列腺癌样本将从NCI申办的ECOG试验中获得;乳腺癌患者和对照样本将由Hoosier肿瘤组专门为此联盟从整个州和地区收集。从印第安纳州大学医学院组织的标准化样本采集将确保患者人口统计学群体的多样性。另外三家公司合作伙伴将提供关键技术和咨询，以提高团队效率。

DESCRIPTION (provided by applicant): Biomarker identification and characterization holds great promise for more precise diagnoses and for tailored therapies. The heterogeneity of human cancers and unmet medical needs in these diseases provides a compelling argument to focus biomarker development in cancer. Mass spectrometry (MS)-based proteomics approaches have provided insight into biomarkers of cancer and other diseases with femtomole sensitivity and high analytical precision, but the results have proven difficult to reproduce owing to the complexity of human biofluids and to the protocols employed in these approaches. This CPTAC proposal will develop robust protocols and standards for MS proteomics employing both electrospray ionization (ESI) and Matrix Assisted Laser Desorption Ionization (MALDI) platforms. Ion mobility MS provides several orders of magnitude increased dynamic range, and protocols for detection of cancer biomarker candidates will be further explored with this emerging technology platform. The consortium will employ high specificity immunologic reagents to develop platforms for precise detection and quantification of biomarkers of relevance for breast and prostate cancer. Immunoaffinity selection of specific biomarker proteins will be achieved as a sample fractionation step using nano-scale immunoaffinity columns prior to MS detection and quantification of specific proteins. An emerging technology incorporating specific antibodies on a microfabricated "bioCD" read by spinning disc interferometry enables label-free evaluation of hundreds of analytes from hundreds of samples in minutes. We will focus on candidate biomarkers relevant for breast and prostate cancer from the NF?B and STATS signaling pathways. Three for-profit and one not-for-profit corporate partners will join several academic groups with deep expertise and extensive resources in proteomics technologies to most efficiently evaluate and roll out robust protocols and standards for MS and affinity proteomics approaches. Prostate cancer samples will be made available from the NCI-sponsored ECOG trial; breast cancer patient and control samples will be collected specifically for this consortium by the Hoosier Oncology Group from throughout the State and region. Standardized sample collection organized from the Indiana University School of Medicine will ensure a diverse cross section of patient demographic groups. Three additional corporate partners will provide key technologies and consultation to improve team efficacy.