Porous Silicon Optical Sensor for Candida Detection

描述（由申请人提供）：本研究计划的主旨是促进对基于多孔硅微腔的相对较新的创新无标签光学生物传感器技术的基本理解。具体目的是研究制造假丝酵母检测亲和传感器的关键参数。噬菌体展示scFv抗体探针，靶向细胞壁表面抗原作为受体。目标将是发现这些探针是否可以在固定化形式中起作用，以区分假丝酵母芽生孢子（非侵入性）和丝状（致病性）形式。通过了解多孔硅微观结构（孔隙度，孔隙形态）和表面化学对生物分子渗透和传感器灵敏度的依赖，将获得基础知识。一个关键的目标将是确定传感器可以检测念珠菌抗原在原发性皮肤感染中典型表达的水平。原发性念珠菌感染的小鼠模型将被用作皮肤抗原的来源。为了实现这些目标，将使用报告分析来量化固定scFv探针相对于溶液控制的活性。初始工作将使用单探针芯片格式的设备完成。罗彻斯特理工学院的硅微加工设备将用于研究制造阵列探针芯片格式所需的工艺步骤，以便同时分析多个目标。访问该设施还将使研究能够阐明对传感器制造的材料科学挑战的基本见解。拟议的研究将产生广泛扩展到其他病原生物和蛋白质组偶联物检测的知识。将获得基本技能，这些技能是成为生物医学科学领域成功的独立研究者的培训过程的关键组成部分。

DESCRIPTION (provided by applicant): The thrust of this research proposal is to advance fundamental understanding of a relatively new and innovative label-free optical biosensor technology based on porous silicon microcavities. The specific aims are designed to investigate key parameters in the fabrication of an affinity sensor for the detecting Candida. Phage display scFv antibody probes that target cell wall surface antigen will serve as the receptor. A goal will be to discover whether these probes can function in an immobilized format to distinguish between the Candida blastoconidia (noninvasive) and filamentous (pathogenic) forms. Foundation knowledge will be gained on understanding the dependence of the porous silicon microstructure (porosity, pore morphology) and surface chemistry on biomoiecular infiltration and sensor sensitivity. A key aim will be to determine the sensor can detect Candida antigen at levels typically expressed in a primary skin infection. A mouse model of primary Candida infection will be utilized as a source of antigen from skin. To accomplish these goals a reporter assay will be used to quantify the activity of the immobilized scFv probes relative to a solution control. Initial work will be done using devices in a single-probe chip format. The silicon microfabrication facility at the Rochester Institute of Technology will be leveraged to investigate the process steps required to fabricate an arrayed-probe chip format so multiple targets could be assayed simultaneously. Access to this facility will also enable studies to elucidate fundamental insight into the material science challenges of sensor fabrication. The proposed research will generate knowledge that is broadly extensible to the detection of other pathogenic organisms and proteomic conjugates. Essential skills will be acquired that comprise a critical component of the training process to become a successful independent investigator in biomedical sciences.