Porous Silicon Optical Sensor for Candida Detection

描述（由申请人提供）：本研究提案的主旨是推进对基于多孔硅微腔的相对新颖和创新的无标记光学生物传感器技术的基本理解。本论文的主要目的是研究亲和传感器制备过程中的关键参数。噬菌体展示靶向细胞壁表面抗原的scFv抗体探针将充当受体。一个目标将是发现这些探针是否可以在一个固定的格式，以区分念珠菌芽生分生孢子（非侵入性）和丝状（致病性）的形式。基础知识将获得理解多孔硅微观结构（孔隙率，孔形态）和表面化学对生物分子渗透和传感器灵敏度的依赖性。一个关键的目标将是确定传感器可以检测念珠菌抗原的水平通常表达在原发性皮肤感染。将使用原发性念珠菌感染的小鼠模型作为皮肤抗原来源。为了实现这些目标，将使用报告基因测定来定量固定的scFv探针相对于溶液对照的活性。最初的工作将使用单探针芯片格式的设备完成。罗切斯特理工学院的硅微加工设施将被用来研究制造阵列探针芯片格式所需的工艺步骤，以便同时测定多个目标。访问该设施还将使研究能够阐明传感器制造的材料科学挑战的基本见解。拟议的研究将产生的知识，是广泛扩展到其他病原体和蛋白质组结合物的检测。将获得基本技能，包括培训过程的关键组成部分，成为一个成功的生物医学科学独立调查员。

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.