CAREER: Understanding Multivalent Biological Bonds for Biosensor Applications

职业：了解生物传感器应用的多价生物键

• 批准号: 1055437
• 负责人: Todd Sulchek
• 金额: $ 40万
• 依托单位: Georgia Tech Research Corporation
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2017-03-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1055437&HistoricalAwards=false
• 关键词: CAREER; Understanding; Multivalent; Biological; Bonds

1055437SulchekCAREER: Understanding Multivalent Biological Bonds for Biosenssing ApplicationsThis project?s research goal is to understand multivalent biological bonds, the simultaneous interaction of multiple ligands with multiple receptors, in order to improve the fields of medicine and bioengineering. Multivalent bonds underpin a variety of biological and biotechnological processes, including cell/tissue adhesion and antibody/therapeutic targeting. However, there is currently no coherent description of multivalent binding which can predict the strength of multivalent interactions and understand the rapid intermediate binding states that may result due to the fluctuations of individual molecular bonds. The research goals will be pursued in a basic science study of the adhesion of a remarkable form of antibody derived from the lamprey eel: the variable lymphocyte receptor. Variable lymphocyte receptor antibodies are highly multivalent and have a well-defined and easily modified structure. Moreover, the antibodies offer several exciting possibilities for biosensor advancement, including high binding affinity with an easy method for varying that affinity as well as incredible environmental stability.The research objectives will be to first create well-defined, multivalent constructs of antibodies. The antibodies will be molecularly engineered to vary the valency and the linker architecture. Second, the bond strength will be measured as valency and linker structure are varied. The bond strength will be measured with a complementary suite of analytical techniques including surface plasmon resonance as well as single-molecule adhesion techniques based upon atomic force microscopy. Force spectroscopy will be used to measure the kinetics of the multi-step multivalent transitions?a capability that is uniquely suited to single-molecule measurements. Third, these measurements will then test the predictions of a new theoretical model describing multivalent binding. The education objectives of this proposal will be to create a cross-disciplinary science fair for an Atlanta public high school; design and implement outreach curricula in an Atlanta elementary school that focus on hands-on science enrichment; and integrate the concepts of biological multivalency into lessons for the Biologically Inspired Design Projects Laboratory at Georgia Tech. Intellectual merit of this research will be to test the hypothesis that multivalent bonds dramatically increase binding strength through a decrease in multi-state dissociation kinetics and that the kinetics of dissociation are radically controlled by adjusting the physical properties and architecture of the linker between multiple antigen binding sites. This research may explain an intriguing question: why have researchers shown such a wide a range of multivalent affinity increase, from 101-1010 fold? The broader impacts of the fundamental understanding of biomolecular adhesion at micro- and nano-interfaces may also improve sensor operation by identifying design principles for highly efficient multivalent binding. Biomolecular interactions can then be tailored to show a wide variety of binding strengths and perhaps sensitivities to near neighbors of a targeted microbial strain. Such capabilities will greatly improve biosensors by reducing the cost and complexity of biosensors while increasing their reliability. Ultimately, this basic science research study will create detection technologies with higher stability, specificity, and sensitivity than can be currently achieved and at the same time provide cross-disciplinary training and education to students at Georgia Tech and local area schools.

这个项目？S的研究目标是了解多价生物键，即多个配体与多个受体同时作用，以促进医学和生物工程领域的发展。多价键支持各种生物和生物技术过程，包括细胞/组织黏附和抗体/治疗靶向。然而，目前还没有关于多价结合的连贯描述，可以预测多价相互作用的强度，并理解由于单个分子键的波动可能导致的快速中间结合状态。这项研究的目标将在一项基础科学研究中进行，该研究旨在研究鳗鱼产生的一种特殊形式的抗体的粘附性：可变淋巴细胞受体。可变淋巴细胞受体抗体是高度多价的，具有明确的和容易修改的结构。此外，抗体为生物传感器的发展提供了几种令人兴奋的可能性，包括高结合亲和力和一种简单的改变亲和力的方法，以及令人难以置信的环境稳定性。研究目标将是首先创造定义明确的多价抗体结构。抗体将进行分子工程，以改变价态和连接物的结构。其次，随着价态和连接基结构的变化，将测量键合强度。结合强度将通过一套互补的分析技术来测量，包括表面等离子激元共振以及基于原子力显微镜的单分子粘合技术。力光谱将被用来测量多步多价转变的动力学--这是一种唯一适合于单分子测量的能力。第三，这些测量将检验描述多价结合的新理论模型的预测。这项提议的教育目标将是为亚特兰大一所公立高中创建一个跨学科的科学博览会；在亚特兰大一所小学设计和实施注重实践科学丰富的推广课程；将生物多价性的概念纳入佐治亚理工学院生物启发设计项目实验室的课程。这项研究的智力价值将检验这样的假设，即多价键通过降低多态解离动力学来显著增加结合强度，并且解离动力学通过调节多个抗原结合位点之间连接物的物理性质和结构来从根本上控制解离动力学。这项研究可能解释了一个有趣的问题：为什么研究人员显示出如此广泛的多价亲和力增加，从101到1010倍？通过确定高效多价结合的设计原则，对微纳界面生物分子黏附的基本了解的更广泛的影响也可能改善传感器的操作。然后，可以定制生物分子相互作用，以显示各种结合强度，或许还可以显示对目标微生物菌株的近邻的敏感性。这种能力将通过降低生物传感器的成本和复杂性，同时提高其可靠性，从而极大地改进生物传感器。最终，这项基础科学研究将创造出比目前能够实现的更高稳定性、特异性和灵敏度的检测技术，同时为佐治亚理工学院和当地学校的学生提供跨学科的培训和教育。