Smart Polymer Reagents for Sensitivity and Speed-enhanced Clinical Diagnostics

描述（由申请人提供）：磁珠作为一种方便的分离技术，越来越多地用于多种形式的细胞、核酸和蛋白质的分离和分析。特别地，临床免疫测定的制造商利用磁珠作为特异性靶向临床重要分析物的抗体的固体支持物，并作为分离和检测那些结合的分析物的分离手段。但是使用磁珠会带来一个矛盾。磁珠必须具有足够的尺寸-通常直径在几微米的量级-以便在容易实现的磁场中分离。但是这种大小的磁珠扩散非常缓慢，与其体积相比，抗体结合的表面积有限。因此，当前磁珠的尺寸限制了临床测定中可以实现的速度和灵敏度。通过模板导向聚合物合成和纳米技术的进步，可以制造出一类新的“智能”磁性纳米颗粒。这些磁性纳米颗粒可以响应于环境刺激（如温度或pH变化）从直径约20纳米的单分散小直径颗粒变化为微米直径的大聚集体。通过使用这些先进的纳米材料，可以开发这样的测定，其中智能磁性纳米颗粒的高表面体积比和小尺寸/高扩散提供了与当前磁珠试剂相比更高的灵敏度和更快的结合反应。然后，离散的pH或温度刺激可以使这些纳米颗粒试剂共聚集成微米尺寸的大聚集体，其可以像目前使用的磁珠一样容易且快速地通过磁场分离。该项目将开发这种”智能”检测试剂，然后在p24（HIV）蛋白免疫测定模型中证明其价值。这些材料有望为心脏病、癌症、内分泌和传染病的重要生物标志物提供更快、更灵敏的临床免疫测定。 公共卫生相关性：存在许多实例，例如甲状腺球蛋白或肌钙蛋白的测定，其中临床测定的灵敏度的增加导致对疾病状态的更丰富的理解和对患者的更好的医疗管理。也有一些例子，如术中促甲状腺激素测定，其中更快的测定导致麻醉时间减少和明显的患者受益。该项目评估了新开发的智能试剂的前景，以实现更灵敏、更快速的临床检测，从而为更好的医疗保健做出贡献。

DESCRIPTION (provided by applicant): Magnetic beads have gained increasing use as a convenient separation technique for many forms of cell, nucleic acid and protein isolations and analyses. In particular, the manufacturers of clinical immunoassays utilize magnetic beads both as a solid support for antibodies specifically targeted to analytes of clinical importance, and as the separation means to isolate and detect those bound analytes. But use of magnetic beads imposes a paradox. The magnetic beads must have sufficient size - normally on the order of several microns in diameter - in order to be separated in an easily achievable magnetic field. But magnetic beads of this size diffuse only very slowly, and present limited surface area for antibody binding compared to their volume. Thus the size of the current magnetic beads limits the speed and sensitivity that can be achieved in clinical assays. Through advances in template-directed polymer synthesis and nanotechnology, a new class of "smart" magnetic nanoparticles can be made. These magnetic nanoparticles can change from a monodispersed small diameter particle of roughly 20 nanometers diameter to a macro- aggregate of microns diameter in response to an environmental stimulus like a temperature or pH change. By using these advanced nanomaterials, assays can be developed in which the high surface to volume ratio and the small size / high diffusion of the smart magnetic nanoparticles provides for higher sensitivity and faster binding reactions compared to current magnetic bead reagents. And then a discrete pH or temperature stimulus can cause these nanoparticle reagents to co-aggregate into a macro-aggregate of micron dimensions which can be separated by a magnetic field as easily and quickly as currently used magnetic beads. This project will develop such" smart" assay reagents and then demonstrate their value in a model p24 (HIV) protein immunoassay. These materials present the promise of faster, more sensitive clinical immunoassays for important biomarkers of cardiac disease, cancer, endocrine and infectious diseases. PUBLIC HEALTH RELEVANCE: There exist many examples, such as assays for thyroglobulin or troponin, where increases in the sensitivity of clinical assays has led to a richer understanding of a disease state and to better medical management of patients. There are also examples, such as intra-operative thyroid stimulating hormone assays, where a faster assay leads to decreased time under anesthesia and obvious patient benefit. This project evaluates the promise of a newly developed smart reagent to enable more sensitive and faster clinical assays, and thus to contribute to better medical care.