APT: the Analytical Proteomics Team

描述(由申请人提供)：生物标记物的识别和表征为更准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中未得到满足的医疗需求为关注癌症的生物标记物开发提供了一个令人信服的论点。基于质谱仪(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洞察，但事实证明，由于人体生物流体的复杂性以及这些方法所采用的方案，结果很难重现。CPTAC的这项提案将为使用电喷雾电离(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的动态范围，利用这一新兴技术平台将进一步探索检测癌症生物标记物候选的协议。该联盟将使用高特异性免疫试剂来开发平台，用于精确检测和量化与乳腺癌和前列腺癌相关的生物标记物。在MS检测和定量特定蛋白质之前，将使用纳米级免疫亲和色谱柱作为样品分级步骤，实现对特定生物标记蛋白质的免疫亲和选择。一项通过旋转圆盘干涉法在微型制造的“生物CD”上结合特定抗体的新兴技术，可以在几分钟内对数百个样本中的数百个分析物进行无标记评估。我们将重点关注与乳腺癌和前列腺癌相关的候选生物标记物，这些标志物来自于NF？B和STATS信号通路。三个营利性和一个非营利性企业合作伙伴将加入在蛋白质组学技术方面拥有深厚专业知识和广泛资源的几个学术小组，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赞助的ECOG试验中获得；乳腺癌患者和对照样本将由Hoosier肿瘤学集团从整个州和地区专门为该财团收集。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。另外三家企业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.