APT: the Analytical Proteomics Team

描述（由申请人提供）：生物标志物的鉴定和表征为更精确的诊断和量身定制的治疗提供了巨大的希望。人类癌症的异质性和这些疾病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）的蛋白质组学方法为癌症和其他疾病的生物标志物提供了具有飞摩尔灵敏度和高分析精度的见解，但由于人体生物流体的复杂性以及这些方法中采用的协议，结果已被证明难以重现。该CPTAC提案将采用电喷雾电离（ESI）和基质辅助激光解吸电离（MALDI）平台，为MS蛋白质组学开发强大的协议和标准。离子迁移率质谱提供了几个数量级的动态范围增加，癌症生物标志物候选物的检测方案将在这个新兴的技术平台上进一步探索。该联盟将采用高特异性免疫试剂来开发与乳腺癌和前列腺癌相关的生物标志物的精确检测和定量平台。特异性生物标记蛋白的免疫亲和选择将在MS检测和定量特异性蛋白之前使用纳米级免疫亲和柱作为样品分离步骤来实现。一项新兴技术将特异性抗体结合到微制造的“生物光盘”上，通过旋转圆盘干涉测量法读取，可以在几分钟内对数百个样品的数百种分析物进行无标签评估。我们将重点关注NF中与乳腺癌和前列腺癌相关的候选生物标志物。B和STATS信号通路。三家营利性企业和一家非营利性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稳健协议和标准。前列腺癌样本将从nci赞助的ECOG试验中获得；乳腺癌患者和对照样本将由Hoosier肿瘤小组专门从全州和地区收集。由印第安纳大学医学院组织的标准化样本收集将确保患者人口统计群体的多样化横截面。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.