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。特别是，临床免疫测定的制造商利用磁珠作为临床重要分析物特异性抗体的坚实支持，并作为分离和检测这些结合分析物的分离手段。但磁珠的使用带来了一个悖论。磁珠必须有足够的尺寸——通常直径为几微米——以便在一个容易达到的磁场中分离。但是这种大小的磁珠扩散非常缓慢，与它们的体积相比，抗体结合的表面积有限。因此，当前磁珠的大小限制了临床检测的速度和灵敏度。通过模板定向聚合物合成和纳米技术的进步，可以制造出一类新的磁性纳米颗粒。这些磁性纳米颗粒，承载刺激响应聚合物，可以从单分散的直径约20纳米的小颗粒转变为直径微米的宏观聚集体，以响应环境刺激，如温度或pH值的变化。通过使用这些先进的纳米材料，可以开发出与目前的磁珠试剂相比，磁性纳米颗粒的高表面积体积比和小尺寸/高扩散提供更高的灵敏度和更快的结合反应的检测方法。然后，离散的pH值或温度刺激可以使这些纳米粒子试剂迅速聚集成微米级的宏观复合物，这种复合物可以像目前使用的磁珠一样，通过磁场轻松快速地分离。该项目将开发、优化并开始扩大这些独特的分析试剂的规模，然后将在p24 （HIV）蛋白模型免疫分析中证明它们的价值。该项目的第二阶段还将展示使用这些试剂进行多种类型艾滋病毒检测的能力，并展示消除某些重要临床分析的血清干扰的潜力。本项目的HIV靶检测只是一个模型。这些材料有望为心脏病、癌症、内分泌失调和其他传染病的重要生物标志物提供更快、更敏感的临床免疫分析。这些试剂的多重功能和去除血清干扰的潜力将广泛适用于其他临床重要分析物。

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 magnetic nanoparticles can be made. These magnetic nanoparticles, bearing stimuli-responsive polymers, 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 area to volume ratio and the small size / high diffusion of the 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 rapidly co-aggregate into a macro-complex of micron dimensions which can be separated by a magnetic field as easily and quickly as currently used magnetic beads. This project will develop, optimize, and begin scale-up of these unique assay reagents, and will then demonstrate their value in a model p24 (HIV) protein immunoassay. Phase II of this project will also demonstrate the ability to multiplex various types of HIV tests using these reagents, and demonstrate the potential of removing serum interferences for certain important clinical analytes. The HIV target assay of this project is only a model. These materials present the promise of faster, more sensitive clinical immunoassays for important biomarkers of cardiac disease, cancer, endocrine disorders and other infectious diseases. The multiplex capabilities of these reagents and the potential for removal of serum interferences will be broadly applicable to other clinically important analytes.