Monoclonal antibody mediated biomarker discovery for alcohol-induced liver damage

单克隆抗体介导的酒精性肝损伤生物标志物的发现

• 批准号: 7414655
• 负责人: LAWRENCE DOUGLAS SNELL
• 金额: $ 24.01万
• 依托单位: LOHOCLA RESEARCH CORPORATION
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-30 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7414655
• 关键词: Affinity; Affinity Chromatography; Alcohol dependence; Alcoholic Liver Diseases; Alcoholic liver damage; Alcohols; Antibodies; Antigens; Biochemical Pathway; Biological; Biological Assay; Biological Markers; Biological Process; Blood; Boston; Classification; Clinical; Clinical Research; Collaborations; Complex; Control Groups; Detection; Development; Diagnostic; Disease; Early Diagnosis; Excision; Gene Expression; Gene Expression Profile; Goals; Hepatocyte; Human; Hybridomas; Immunoassay; Immunology; Individual; Institutes; International; Libraries; Mass Chromatography; Mass Spectrum Analysis; Mediating; Methodology; Methods; Molecular; Monoclonal Antibodies; Organ; Outcome; Participant; Pathway interactions; Patients; Pattern; Phage Display; Pharmaceutical Preparations; Phase; Plasma; Plasma Proteins; Polymerase Chain Reaction; Post-Translational Protein Processing; Process; Production; Protein Microchips; Proteins; Proteome; Proteomics; Purpose; Range; Reagent; Relative (related person); Reporting; Research; Sampling; Science; Screening Result; Small Business Funding Mechanisms; Small Business Innovation Research Grant; Specificity; Spectrometry; Staging; Statistically Significant; Systems Biology; Techniques; Technology; Testing; Today; Universities; Validation; alcohol exposure; analytical method; assay development; base; cost; day; interest; liquid chromatography mass spectrometry; liver biopsy; new technology; novel; novel strategies; programs; protein expression; prototype; response; success; technology development; three dimensional structure

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.

描述(由申请人提供)：这项SBIR申请的目标是证明一种全球方法的可行性，以筛选针对低水平蛋白质的疾病特异性单抗(MAbs)，以发现和验证酒精诱导的肝损伤的生物标记物。这项技术将利用与今天遵循的工作流程基本上相反的工作流程，即首先识别抗原，然后为潜在的生物标记物产生抗体。在目前的方案中，将从1000多株杂交瘤中筛选酒精性肝损伤患者的血浆蛋白质组和合适的对照组，以识别抗体，寻找至少10种明显的疾病特异性识别生物标志物单抗。为了证明单抗是低水平的蛋白，将通过亲和分离和质谱仪鉴定至少5个单抗的抗原。血液中这些蛋白质的水平也将被测定或估计。该方法代表了一种发现和验证生物标志物的新方法，与目前基于LC/MS的方法相比，其灵敏度提高了10倍以上，成本降低了10倍，吞吐量提高了许多倍。一旦该方法的可行性得到证明，该方法将准备好在个别患者样本上实施。该项目成功的重要因素是国际生物系统公司与位于马萨诸塞州波士顿的东北大学巴尼特研究所的合作，前者是一家在免疫学和化验开发方面具有专长的初创公司，后者是一家在层析和质谱学方面具有专长的研究中心。当今迫切需要的是开发一种技术，能够快速且经济高效地提供针对血液中低水平蛋白质的疾病特异性抗体，并可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记物。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。