SBIR Phase I: Binding Protein Patterns on Plastic Surfaces

*9661439 Guire本项目旨在展示在特定结合分析和生物传感器系统中使用的塑料材料涂层的普遍适用、简便和经济高效的方法的可行性。用生物分子相容的稳定剂对表面进行共价钝化、生物分子的图案化固定，这些都是生物传感器技术商业化发展的关键和持久的障碍，将通过制备具有光反应性的“万能结合”薄膜表面来解决。在温和的反应条件下，高亲和力的总配体(生物素)衍生物和钝化/稳定剂的共价光化学键将分别通过图案化和均匀光照快速完成。固定在钝化光学级塑料上的X-亲和素的图案化阵列将通过用该多位点结合蛋白的粗溶液或纯化溶液浸涂该表面来获得，然后通过通过喷墨打印(例如，利用针对囊性纤维化基因缺陷的所需生物素化寡核苷酸)不同地加载阵列的单个测定点(例如，利用所需的生物素化寡核苷酸)来制备用于多分析分析的基本上任何核酸探针或特定分析物结合蛋白(例如抗体)的分析。这种方法有望为生物分子-传感器接口的突出技术问题提供具有成本效益的解决方案，这些接口是正在开发的小样本、灵敏、多羟基、坚固耐用的便携式商业设备，用于对环境、食品、传染病和基因序列分析和监测有价值的核酸探针和免疫分析。***

*** 9661439 Guire This project is designed to demonstrate the feasibility of a generally applicable, facile, and cost-effective approach to coating of plastic materials used in specific binding assay and biosensor systems. Covalent passivation of the surface with a biomolecule-compatible stabilizing agent, patterned immobilization of biomolecules, which are critical and enduring obstacles to the commercial development of biosensor technology, will be resolved through the preparation of a photoreactive "universal binding" thin film surface. Covalent photochemical bonding of the high affinity general ligand (biotin) derivative and of the passivating/stabilizing agent, will be accomplished quickly under mild reaction conditions by patterned and uniform illumination, respectively. A patterned array of X-avidin immobilized on passivated optical grade plastic will be obtained by dip-coating this surface with a crude or purified solution of this multi-site binding protein, then prepared for multianalyte assay use with essentially any nucleic acid probes or specific analyte binding proteins (e.g., antibodies) assay through differential loading of the individual assay dots of the array by inkjet printing (e.g., with the desired biotinylated oligonucleotides for cystic fibrosis gene defects). This approach is expected to provide cost-effective solutions to the outstanding technical problems of the biomolecule-sensor interface of small sample, sensitive , multiananyte, rugged portable commercial devices under development for nucleic acid probe and immune assays of value for environmental, food, infectious disease, and gene sequence analysis and monitoring. ***