CAREER: Understanding Multivalent Biological Bonds for Biosensor Applications

1055437 SulchekCAREER：了解生物传感应用中的多价生物键这个项目？的研究目标是了解多价生物键，多个配体与多个受体的同时相互作用，以改善医学和生物工程领域。多价键支持各种生物和生物技术过程，包括细胞/组织粘附和抗体/治疗靶向。然而，目前没有连贯的描述多价结合，可以预测多价相互作用的强度，并了解快速的中间结合状态，可能会导致由于个别分子键的波动。研究目标将在一项基础科学研究中进行，该研究是关于来自七鳃鳗的一种显着形式的抗体的粘附：可变淋巴细胞受体。可变淋巴细胞受体抗体是高度多价的，并且具有明确定义且易于修饰的结构。此外，这些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改变亲和力的简单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建定义明确的多价抗体构建体。抗体将被分子工程化以改变化合价和接头结构。第二，随着化合价和连接基结构的变化，将测量结合强度。结合强度将用一套互补的分析技术来测量，包括表面等离子体共振以及基于原子力显微镜的单分子粘附技术。力谱将用于测量多步多价跃迁的动力学？这是一种特别适合于单分子测量的能力。第三，这些测量将测试描述多价结合的新理论模型的预测。该提案的教育目标将是为亚特兰大公立高中创建一个跨学科的科学博览会;在亚特兰大小学设计和实施外展课程，重点是实践科学丰富;并将生物多价的概念融入格鲁吉亚理工学院生物启发设计项目实验室的课程中。本研究的智力价值将是检验以下假设：多价键通过降低多态解离动力学显著增加结合强度，并且解离动力学通过调节多个抗原结合位点之间的接头的物理性质和结构而从根本上控制。这项研究可以解释一个有趣的问题：为什么研究人员显示出如此广泛的多价亲和力增加，从101-1010倍？ 在微米和纳米界面的生物分子粘附的基本理解的更广泛的影响，也可以通过确定高效的多价结合的设计原则，提高传感器的操作。然后可以定制生物分子相互作用，以显示各种各样的结合强度，并可能对目标微生物菌株的近邻敏感。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，大大改善生物传感器。最终，这项基础科学研究将创造比目前更高的稳定性，特异性和灵敏度的检测技术，同时为格鲁吉亚理工学院和当地学校的学生提供跨学科的培训和教育。

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.