Capillary Electrophoresis Profiling of Tears in Dry Eye

描述：（申请人的摘要）我们提出研究和开发一种 廉价和可靠的方法来确定脂质和蛋白质的组成 眼泪可以在基础科学实验室和临床上使用， 在诊断实验室环境中。我们的方法不仅会产生 更精确地诊断干眼症所需的详细信息， 也为制定更具体、更有效的 针对个体的类别（以及可能直接针对个体的治疗） 这些缺陷是干眼症的特征。超过 仅在美国就有1200万人被临床诊断为 患有某种形式的干燥性角膜炎或干眼。虽然他们的症状是 同样，根本原因往往是未知的，也没有实际的方法 用于评估来自这些患者的眼泪的组成。到目前为止， 由于样本量小， 现有的和技术上无法确定和衡量的组成部分， 这种复杂的流体在纳摩尔或更小的浓度尺度。 电喷雾离子化质谱法（CE-ESI/MS）是一种 灵敏、相对便宜的方法，能够分析样品2 μ l或 尺寸较小。采用胶束电动毛细管色谱技术 结合质谱检测，我们计划设计和开发1） 泪液中脂质的新标准分离方法和2）新的 泪液中蛋白质的标准分离方法。然后我们将 研究使用这些方法来开发标准脂质和蛋白质 正常泪液的成分谱，并确定特定成分差异 在两个不同的干眼症模型的眼泪中。发展这个 这项技术提供了关于称为干燥的复杂疾病的具体信息， 眼睛可以提供救济和提高生活质量的数百万人 目前尚无可靠的长效治疗的患者。

DESCRIPTION: (Applicant's Abstract) We propose to investigate and develop an inexpensive and reliable method to determine the lipid and protein composition of tears that could be used both in the basic science laboratory and clinically in a diagnostic laboratory setting. Our approach would not only generate the detailed information needed for more precise diagnosis of dry eye conditions, but also provide the basis for development of more specific and effective categories for (and possibly therapies targeted directly to) the individual deficiencies that ' characterize the spectrum of dry eye disorders. More than 12 million people in the United States alone have been clinically diagnosed with some form of keratitis sicca or dry eyes. Although their symptoms are similar, the underlying causes are often unknown and no practical methodology for evaluating the composition of tears from such patients exists. To date, analysis of tear composition has been hampered by the minute sample sizes available and the technical inability to identify and measure the components of such a complex fluid at concentration scales of nanomoles or less. Capillary electrophoresis-electrospray ionization mass spectrometry (CE-ESI/MS) is a sensitive, relatively inexpensive method capable of analyzing samples 2 pl or less in size. Using micellar electrokinetic capillary chromatography techniques coupled with mass spectrometry detection, we plan to design and develop 1) a novel standard separation method for the lipids within tears and 2) a novel standard separation method for the proteins within tears. We will then investigate the use of these methods to develop standard lipid and protein component profiles of normal tears and identify specific component differences in the tears from two different dry eye models. The development of this technology to provide specific information on the complex disorder know as dry eye could provide relief to and enhance the quality of life of millions of patients for whom no reliable long-lasting therapy is currently available.