NOVEL METHOD TO MEASURE GLOMERULAR SIZE-SELECTIVITY

我们的具体目标是验证和进一步开发一种创新的方法来测量肾小球的大小选择性。我们建议使用一系列新颖的标记，预尺寸的惰性聚合物。通过这种方法，可以直接从收集的血清和尿液样本中获得肾小球的大小选择性测量。这种方法将消除与放射性相关的缺点和繁琐的步骤，即通过尺寸专用色谱法分离大分子片段。该方法将提供一种安全、易于使用、准确且具有成本效益的肾小球大小选择性检测试剂盒，同时为广泛的研究和临床情况提供一种通用的诊断方式。实验将描述我们的新标签系统。标记的聚合物将被测试每个聚合物的标签键数；总体变化;温度变化对缓冲液孵育的稳定性；稳定性挑战与竞争的螯合剂和在血清环境。将确定最佳贴标条件以及检测灵敏度。

Our specific aim is to validate and further develop an innovative method to measure glomerular size-selectivity. We propose to use a series of novelly labeled, pre-sized inert polymers. By this method, glomerular size-selectivity measurements can be directly obtained from collected serum and urine samples. This approach will eliminate both the disadvantages associated with radioactivity and the cumbersome step of post-separating macromolecules fragments by size-exclusive chromatography. This approach will provide a glomerular size-selectivity assay kit that is safe, easy-to-use, accurate, and cost-effective while providing a diagnostic modality that is versatile for a wide range of research and clinical situations. Experiments will characterize our novel labeling system. Labeled polymer will be tested for the number of labels bond per polymer; overall change; stability to incubation in buffer with temperature variation; stability to challenge with competing chelates and in a serum environment. The optimum labeling conditions will be defined as well as the sensitivity of detection. PROPOSED COMMERCIAL APPLICATION: Research is directed toward the development of an easy-to-use and cost-effective glomerular size-selectivity assay kit, suitable for both the research and clinical marketplace. This product will provide increased assay design flexibility over current methods.