An Enzyme Linked Immunosorbent Assay (ELISA) for Monitoring Galactosemia

描述(由申请人提供)：这项提案的长期目标是开发一种诊断试剂盒，用于监测半乳糖中的非酶半乳蛋白。经典的半乳糖是由于缺乏半乳糖-1-磷酸尿苷转移酶而引起的半乳糖代谢紊乱。由此造成的严重的半乳糖代谢障碍已经被认识到半个世纪了，这种疾病的频率估计是每40,000名新生儿中就有1名。通常，生化表型包括组织和体液中半乳糖、半乳糖-L-磷酸和半乳糖醇浓度升高。虽然这种疾病通常是致命的，但严格限制饮食半乳糖的治疗已被证明是挽救生命的；然而，尽管这样饮食，大多数患者仍会出现神经异常。早期的研究表明，半乳糖血症患者糖化血红蛋白/白蛋白水平升高，这促使人们推测，类似于糖化蛋白糖尿病，监测半乳糖化蛋白可能被证明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半乳糖化白蛋白的单抗探针，通过标准的免疫学技术将证明它是针对半乳糖化白蛋白的。该探针将用于酶联免疫吸附试验，以确定其灵敏度和检测范围。第二阶段的工作将集中于在临床环境中评估这种酶联免疫吸附试验，并优化其用于治疗半乳糖血症患者的性能。第三阶段的工作将集中在这种检测的商业化上。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长期以来一直在等待监测半乳糖血症患者的检测方法；对半乳糖蛋白进行量化的方法可能为预防或推迟这种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出现神经损伤。

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.