Suspension arrays detection/quantification of antibodies

描述(申请人提供)：在现代医学和科学中，越来越需要能够分析血清和唾液等体液的成分。微阵列芯片技术具有提供这种能力的潜力。这项技术已经改变了制药研究。然而，到目前为止，这项技术还没有被广泛应用于以蛋白质为重点的研究。其原因在于蛋白质性质的巨大多样性。这种多样性影响蛋白质阵列的性能，因此限制了这种阵列可以同时查询的分析物的数量。我们的目标是开发一套相互关联的协议和结构，以构成蛋白质阵列设计的新方法。这种方法将实现更统一的效率：1)将探针蛋白质附着到阵列的基质上，2)单个探针与来自询问样本(分析物)的相应蛋白质之间的相互作用，以及3)单个相互作用对的可视化。为了实现我们的目标，我们将把配体展示技术和嵌合体蛋白产生技术的进步与新兴的基于颗粒的流式细胞仪分析平台结合起来。首先，我们打算应用已开发的方法来满足生物恐怖主义防御领域的现有需求。因此，在第一阶段，我们将通过开发一种检测A型肉毒杆菌血清抗体的阵列来证明该方法的可行性。在第二阶段，我们将扩大该系统检测到的抗体的范围。我们将把炭疽芽孢杆菌、肉毒梭菌(B、C和E型)和蓖麻毒素等生物恐怖主义领域令人担忧的病原体的抗原决定因素纳入阵列，以创建一个工具来评估个人免受潜在生物恐怖袭击的保护水平。

DESCRIPTION (provided by applicant): In modern medicine and science there is a growing need for the ability to profile composition of such bodily fluids as serum and saliva. Microarray chip technology has potential for delivering such ability. This technology has already changed pharmaceutical research. So far, however, this technology has not been used extensively in protein-focused studies. The reason for this lies in the substantial diversity of protein properties. This diversity affects performance of protein arrays and, consequently, limits the number of analytes that can be simultaneously interrogated by such arrays. Our objective is to develop a set of interconnected protocols and constructs that will constitute a novel approach to protein array design. This approach will allow achieving more uniform efficiencies of: 1) attachment of probe proteins to the matrix of the array, 2) interaction between individual probes and corresponding proteins from the interrogating sample (analytes), and 3) visualization of individual interacting pairs. To achieve our objective we will combine advancements in the ligand-display technology and techniques for generation of chimera proteins with the emerging platform of particle-based flow cytometric assays. First, we intend to apply the developed approach to address the needs existing in the area of bioterrorism defense. Therefore, during Phase I we will demonstrate the feasibility of the approach by developing an array for detection of anti-botulinum serotype A antibodies. During Phase II, we will broaden the spectrum of antibodies detected by the system. We will incorporate into the array antigenic determinants for such pathogenic agents of concern in the field of bioterrorism as Bacillus anthracis, Clostridium botulinum (serotypes B, C and E), and ricin to create a tool for evaluation of the level of individual protection from potential bioterrorist attacks.