Monoclonal antibody mediated biomarker discovery for alcohol-induced liver damage

描述（由申请人提供）：本SBIR申请的目的是证明筛选低水平蛋白质的疾病特异性单克隆抗体（mAb）的全球方法的可行性，以发现和验证酒精诱导肝损伤的生物标志物。该技术将利用一个基本上与今天相反的工作流程，首先识别抗原，然后为潜在的生物标志物生成抗体。在本提案中，酒精诱导的肝损伤患者的血浆蛋白质组和合适的对照组将针对超过1000个杂交瘤筛选用于区分抗体，寻求至少10种明显疾病特异性区分生物标志物mAb。为了证明mAb是低水平蛋白，将通过亲和分离和质谱法鉴定至少5种mAb的抗原。还将确定或估计血液中这些蛋白质的水平。所建议的方法代表了一种用于生物标志物发现和验证的新方法，提供了超过10倍的灵敏度，以及比目前基于LC/MS的方法便宜10倍和高出许多倍的通量。一旦该方法的可行性得到证明，该方法将准备在个体患者样本上实施。该项目的成功离不开Biosystems International（一家在免疫学和检测开发方面拥有专业知识的初创公司）与位于马萨诸塞州波士顿的东北大学巴内特研究所（一家在色谱和质谱方面拥有专业知识的研究中心）的合作。当今的一个关键需求是开发一种技术，该技术可以快速且具有成本效益地提供针对血液中低水平蛋白质的疾病特异性抗体，并且可以用于发现和验证用于早期检测酒精诱导的肝损伤的生物标志物。该计划的目标是满足这一需求，所产生的试剂应广泛用于诊断目的。

DESCRIPTION (provided by applicant): The goal of this SBIR application is to demonstrate the feasibility of a global approach to screen for disease specific monoclonal antibodies (mAbs) to low level proteins for the discovery and validation of biomarkers of alcohol-induced liver damage. The technology will utilize a workflow that is essentially the reverse of that followed today, where antigens are first identified and then antibodies generated for potential biomarkers. In the present proposal, plasma proteome of alcohol-induced liver damage patients and a suitable control group will be screened against more than 1000 hybridomas for discriminating antibodies, seeking for at least 10 apparently disease specific discriminating biomarker mAbs. To demonstrate that the mAbs are to low level proteins, the antigens for at least 5 mAbs will be identified by affinity isolation and mass spectrometry. The level of these proteins in blood will also be determined or estimated. The suggested approach represents a novel method for biomarker discovery and validation, offering more than 10-fold greater sensitivity, as well as being 10-fold less expensive and many-fold higher throughput than present day LC/MS based approaches. Once feasibility of the approach has been demonstrated, the methodology will be ready for implementation on individual patient samples. Important to the success of this project is the collaboration of Biosystems International, a startup company with expertise in immunology and assay development, with the Barnett Institute at Northeastern University in Boston, MA, a research center with expertise in chromatography and mass spectrometry. A critical need today is the development of technology that can rapidly and cost effectively deliver disease specific antibodies for low level proteins in blood and that can be used to discover and validate biomarkers for the early detection of alcohol-induced liver damage. The goal of this program is to meet this need, and the reagents generated should be widely utilized for diagnostic purposes.