Biomolecular Analysis using Liquid Crystals

使用液晶进行生物分子分析

• 批准号: 6945828
• 负责人: NICHOLAS L ABBOTT
• 金额: $ 46.36万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-03 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6945828
• 关键词: antineoplastics; biochemistry; biological signal transduction; carcinogenesis; chemical kinetics; enzyme linked immunosorbent assay; epidermal growth factor; gene expression; growth factor receptors; high throughput technology; intermolecular interaction; liquid crystal; surface plasmon resonance; technology /technique development; tissue /cell culture; western blottings

DESCRIPTION (provided by applicant): The University of Wisconsin Bioengineering Research Partnership (UW-BRP) will focus on the development of new molecular analysis tools that possess the potential to be used to identify and validate biological endpoints whereby the activity of novel anti-cancer agents can be more accurately and rapidly evaluated as to their molecular mechanism(s) and clinical relevance. The work will initially be focused on the analysis of the epidermal growth factor receptor (EGFR), given that its over expression and mutation has been well-associated with some of the most incurable cancers. However, it is anticipated that the principles to be developed will be sufficiently versatile to be applied to other key signaling molecules in the future. Whereas the basis of existing high throughput screens largely restricts their application to in vitro molecular analyses of enriched preparations of receptors or other signaling molecules, the UW-BRP seeks to establish principles for tools that can also be applied to the analysis of samples from cultured cells and from biopsies of xenographs and spontaneous tumor tissues. This capability will ultimately enable a fundamental approach that will span the molecular, cellular and tissue levels and will be used in both basic research and in animal and human clinical trials. In the present proposal, a multidisciplinary team of researchers with diverse expertise in chemical and biological engineering, chemistry and biochemistry, and the biomolecular and biomedical sciences proposes to develop a broadly-applicable bioanalytical approach that integrates advances in the following areas: a) the nano-fabrication of surfaces, b) the development of synthetic and biochemical strategies for the covalent and oriented immobilization of proteins and peptides on surfaces, c) the implementation of liquid crystals as highly sensitive reporters of the presence of proteins captured on surfaces, and d) the investigation of key cell signaling proteins that participate in processes associated with carcinogenesis. Specifically, the analytical characteristics of liquid crystals for reporting the behavior of the well-recognized anti-cancer target, i.e. the EGF receptor, will be compared to conventional analytical methods in a study that will a) rapidly and sensitively assess the levels and activity of wild-type and mutant human EGF receptor in biological samples, b) test the hypothesis that wild-type and oncogenic forms of the EGF receptor exhibit differential inhibitor specificity, and c) assess if agents that potently inhibit EGF-mediated events in vitro will also exhibit a capacity to antagonize EGF receptor expression and/or activity in cell culture. These studies will use the EGF receptor system as a prototype and it is anticipated that the technology will be readily adaptable to a wide range of other molecular targets. In the long term, these new tools should be useful for the assessment of the molecular mechanisms and consequences of anti-cancer agents, thereby facilitating their research from basic biology through to clinical assessment of efficacy.

描述（由申请人提供）：威斯康星大学生物工程研究合作伙伴（UW-BRP）将重点开发新的分子分析工具，这些工具有潜力用于识别和验证生物终点，从而确定新型抗癌药物的活性可以更准确、更快速地评估药物的分子机制和临床相关性。鉴于表皮生长因子受体（EGFR）的过度表达和突变与一些最无法治愈的癌症密切相关，这项工作最初将集中于表皮生长因子受体（EGFR）的分析。然而，预计要开发的原理将具有足够的通用性，可以在未来应用于其他关键信号分子。尽管现有高通量筛选的基础在很大程度上限制了其在受体或其他信号分子富集制剂的体外分子分析中的应用，但 UW-BRP 寻求建立也可应用于分析培养细胞样品的工具原理以及异种移植和自发肿瘤组织的活检。这种能力最终将实现一种跨越分子、细胞和组织水平的基本方法，并将用于基础研究以及动物和人类临床试验。在本提案中，由在化学和生物工程、化学和生物化学以及生物分子和生物医学领域拥有不同专业知识的多学科研究人员团队提议开发一种广泛适用的生物分析方法，该方法整合了以下领域的进步： -表面的制造，b) 开发用于将蛋白质和肽共价和定向固定在表面上的合成和生化策略，c) 使用液晶作为存在的高度灵敏的报告剂表面捕获的蛋白质，d) 研究参与癌发生相关过程的关键细胞信号蛋白。具体来说，用于报告公认的抗癌靶标（即 EGF 受体）行为的液晶分析特性将与研究中的传统分析方法进行比较，该研究将：a）快速、灵敏地评估生物样品中的野生型和突变型人类 EGF 受体，b) 测试 EGF 受体的野生型和致癌形式表现出不同的抑制剂特异性的假设，以及 c) 评估在体外有效抑制 EGF 介导事件的药物是否会还表现出拮抗细胞培养物中EGF受体表达和/或活性的能力。这些研究将使用 EGF 受体系统作为原型，预计该技术将很容易适应广泛的其他分子靶点。从长远来看，这些新工具应该有助于评估抗癌药物的分子机制和后果，从而促进从基础生物学到临床疗效评估的研究。