Smart Polymer Reagents for Sensitivity and Speed-enhanced Clinical Diagnostics

描述(申请人提供)：磁珠作为一种方便的分离技术越来越多地被用于多种形式的细胞、核酸和蛋白质的分离和分析。特别是，临床免疫分析的制造商利用磁珠作为针对具有临床重要性的分析物的抗体的固体载体，以及作为分离手段来分离和检测那些结合的分析物。但磁珠的使用带来了一个悖论。磁珠必须有足够的尺寸--通常直径在几微米左右--以便在容易获得的磁场中分离。但这种大小的磁珠扩散速度非常慢，与其体积相比，抗体结合的表面积有限。因此，电流磁珠的大小限制了在临床检测中可以达到的速度和灵敏度。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。这些携带刺激响应性聚合物的磁性纳米颗粒可以随着温度或pH变化等环境刺激而从直径约20纳米的单分散小颗粒转变为直径为微米的大颗粒。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，高比表面积与体积比和小尺寸/高扩散的磁性纳米颗粒提供更高的灵敏度和更快的结合反应的分析方法。然后，离散的pH或温度刺激可以使这些纳米粒子试剂快速共聚集成微米尺度的大分子络合物，它可以像目前使用的磁珠一样容易和快速地被磁场分离。该项目将开发、优化并开始放大这些独特的分析试剂，然后将在模型p24(HIV)蛋白质免疫分析中展示它们的价值。该项目的第二阶段还将展示使用这些试剂对各种类型的艾滋病毒检测进行多重检测的能力，并展示消除某些重要临床分析的血清干扰的潜力。该项目的HIV靶向检测只是一个模型。这些材料有望对心脏疾病、癌症、内分泌紊乱和其他传染病的重要生物标志物进行更快、更灵敏的临床免疫分析。这些试剂的多重能力和消除血清干扰的潜力将广泛适用于其他临床重要的分析方法。

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.