An Enzyme Linked Immunosorbent Assay (ELISA) for Monitoring Galactosemia

描述（由申请人提供）：本提案的长期目标是开发一种用于监测半乳糖中非酶促半乳糖蛋白的诊断试剂盒。经典半乳糖是一种由半乳糖-1-磷酸尿苷转移酶缺乏引起的半乳糖代谢紊乱。由此导致的半乳糖代谢严重受损已经被认识了半个世纪，这种疾病的发病率估计为4万分之一。通常，生化表型包括组织和体液中半乳糖、半乳糖- 1 -磷酸和半乳糖醇浓度升高。虽然这种疾病通常是致命的，但事实证明，严格限制饮食中半乳糖的治疗可以挽救生命；然而，尽管如此，大多数患者还是出现了神经系统异常。早期的研究表明，半乳糖血症中糖化血红蛋白/白蛋白水平升高，这促使人们推测，与糖化蛋白的糖尿病类似，监测半乳糖蛋白可能对治疗半乳糖血症患者有用。在第一阶段，我们将尝试开发一种针对非酶促半乳糖白蛋白的单克隆抗体探针，通过标准免疫学技术将证明该探针对半乳糖白蛋白具有特异性。然后将该探针用于ELISA，以确定其灵敏度和检测范围。II期工作将侧重于在临床环境中评估该ELISA，并优化其管理半乳糖血症患者的性能。第三阶段的工作将侧重于该检测的商业化。Vandalia Research, Inc.致力于制造、营销和销售这项研究成果所产生的诊断试剂盒。儿科医生和内科医生一直在等待检测泌乳症患者；一种量化乳酸蛋白的方法可能为预防或延缓该疾病的许多并发症提供一种可行的方法，包括在儿童期和青春期后发生神经损伤。

DESCRIPTION (provided by applicant): The long term objective of this proposal is to develop a diagnostic kit for monitoring nonenzymatically galactated proteins in galactose. Classical galactose is a disorder of galactose metabolism caused by a deficiency of galactose-1-phosphate uridyltransferase. The resulting severe impairment of galactose metabolism has been recognized for half a century and the frequency of this disorder is estimated to be 1 in 40,000 births. Generally, the biochemical phenotype includes elevated concentrations of galactose, galactose-l- phosphate and galactitol in tissue and body fluids. While the disease is generally fatal, treatment consisting of severe restriction of dietary galactose has proved life saving; nevertheless, most patients develop neurological abnormalities despite this diet. Earlier studies demonstrated the occurrence of elevated levels of glycated hemoglobin/albumin in galactosemia prompting the speculation that, akin to diabetes with glycated proteins, the monitoring of galactated proteins may prove useful for managing galactosemic patients. During Phase I, we will attempt to develop a monoclonal antibody probe directed to nonenzymatically galactated albumin that by standard immunological techniques will prove specific to galactated albumin. This probe will be then used in ELISA to establish its sensitivity and range of detection. Phase II efforts will focus on evaluating this ELISA in a clinical setting and optimizing its performance for the management of galactosemic patients. Phase III efforts will focus on the commercialization of this assay. Vandalia Research, Inc. is committed to manufacture, market and sell the diagnostic kit that will result from this research effort. Pediatricians and internists have long been waiting for assays to monitor galactosemic patients; a method to quantify galactated proteins may provide a viable approach for preventing or delaying many of the complications of the disease including the onset of neurological damage during childhood and post adolescent years.