Cell Mimic Microarrays for Multivalent Pathogen Characterization & Detection

描述（由申请人提供）：本研究项目旨在开发用于病原体分析、表征和检测的细胞模拟微阵列。所提出的方法利用了病原体攻击的初始阶段的共同机制，即通过病原体上的受体蛋白与细胞表面上的碳水化合物（聚糖）分子之间的多价相互作用识别并附着到宿主细胞上。与高度特异性的蛋白质-蛋白质相互作用不同，蛋白质受体与单个聚糖分子之间的低且变化的亲和力通过存在多种相互作用来补偿。有人认为，微阵列表面上显示高密度的聚糖分子可以促进这种多价相互作用。碳水化合物微阵列已被成功地证明在多价结合，包括病原体的检测。尽管取得了初步的成功，一个显着的局限性仍然存在：大多数碳水化合物微阵列证明，迄今为止使用碳水化合物分子共价连接到固体表面。缺乏流动性不能模拟体内细胞表面过程，其中与糖脂和糖蛋白相关的聚糖基团处于流体脂质双层环境中。事实上，认为迁移率是介导多价相互作用的重要因素，例如，在宿主细胞表面聚糖配体的动态聚集中。这是SBIR建议的目的是开发一个平台，碳水化合物微阵列的基础上，专有的空气稳定的支持脂质双层具有高水平的流动性。与其他碳水化合物微阵列相比，所提出的流体阵列更紧密地模仿细胞表面环境，并且可以更有效地应用于病原体吸附的研究。具体目标是制造流体碳水化合物微阵列的基础上纳入空气稳定的支持脂质双层的乙二醇脂质和使用植物凝集素刀豆蛋白A和霍乱毒素B-亚基（CTB）的概念验证实验。一个长期成果将是开发有效的工具，以了解和检测病原体，并制定治疗和预防措施。 该研究项目旨在开发用于理解和检测病原体的细胞模拟微阵列，以及用于预防和治疗病原体攻击的发展。

DESCRIPTION (provided by applicant): This research project aims to develop cell mimic microarrays for the profiling, characterization, and detection of pathogens. The proposed approach exploits a common mechanism at the initial stage of pathogen attack, namely the recognition of and attachment onto host cells via multivalent interaction between receptor proteins on pathogens and carbohydrate (glycan) molecules on cell surfaces. Unlike the highly specific protein-protein interaction, the low and varying affinity between a protein receptor and a single glycan molecule is compensated for by the presence of multiple interactions. It has been argued that the display of a high density of glycan molecules on the surface of a microarray can facilitate such multivalent interaction. Carbohydrate microarrays have been successfully demonstrated in multivalent binding, including the detection of pathogens. Despite the initial successes, a significant limitation remains: most carbohydrate microarrays demonstrated to date use carbohydrate molecules covalently attached to solid surfaces. The lack of mobility does not mimic cell surface processes in vivo where glycan groups associated with glycolipids and glycoproteins are in a fluidic lipid bilayer environment. Indeed, mobility is believed to be a significant factor in mediating multivalent interactions, e.g., in the dynamic clustering of glycan ligands on the host cell surface. It is the purpose of this SBIR proposal to develop a platform for carbohydrate microarrays based on a proprietary air-stable supported lipid bilayer possessing a high level of fluidity. Compared to other carbohydrate microarrays, the proposed fluidic array more closely mimics the cell surface environment and can be applied more efficiently in the study of pathogen adsorption. The specific aims are to fabricate fluidic carbohydrate microarrays based on glycol lipids incorporated into the air-stable supported lipid bilayers and to use plant lectin ConA and cholera toxin B-subunits (CTB) in proof-of-concept experiments. A long-term outcome will be the development of effective tools for the understanding and detection of pathogens, as well as for the development of treatment and prevention. This research project aims to develop cell mimic microarrays for the understanding and detection of pathogens, as well as for the development of prevention & treatment of pathogen attack.