Blood test to aid treatment decisions for perinatal asphyxiation

这种工业-学术合作旨在分析验证一种高度特异性和敏感性的 血清和/或血浆中胶质细胞酸性蛋白（GFAP）和tau的多重免疫测定，以改善 新生儿缺氧缺血性脑病（HIE）的治疗决策。HIE，影响 大约每1 000个新生儿中就有1个是儿童死亡和残疾的主要原因。提示全身 在某些情况下，新生儿冷却可改善结局;然而，24%的冷却新生儿死于HIE， 19%的幸存者发展脑瘫，尽管最好的治疗。低温症的辅助治疗 扩大治疗的有效性。这些研究性佐剂的临床试验进展缓慢， 难以确定哪些新生儿最有可能从治疗中受益。通过预先筛选新生儿 在损伤谱的极端情况下，根据以下因素对新生儿进行客观简单的血液测试 损伤严重程度和监测治疗反应将加速研究性佐剂的试验。 多路复用器采用了一种新的格式，灵敏度比常规格式高出约100-1，000倍 免疫测定，其改善了这些低丰度的临床相关范围内的定量 生物标志物。我们的初步数据表明，我们的新检测格式是足够敏感的测量， 血清和血浆中的GFAP和tau。约翰霍普金斯大学的合作者（埃弗雷特和诺辛顿博士） 已经证明了GFAP和tau作为损伤严重程度和功能结果的标志物的临床实用性， 嗨。该提案旨在充分开发和优化超灵敏多路复用使用我们的相位门控 过程，然后进行广泛的分析验证。 第一年的任务是对关键的关键试剂进行广泛的表征，检测微调，并建立 根据HIE严重程度对新生儿进行分层的关键浓度的分析对照。风险分析 将进行，并将评估分析前因素和潜在干扰物的影响， 减轻。在第二年，设计将被冻结并转移到制造。关键的QC方案 将建立和执行试剂，并根据MSD的ISO配置组件 9001：2015-质量管理体系认证。将在最终设计时生产三个试剂盒批次。测定 将在整个项目中评估性能，包括临床样本的测试。正式 将与临床化学家（JHU的Sokoll博士）合作开发分析试验验证计划， 根据NIH项目官员的意见，并基于2018年FDA生物分析方法验证 工业指导。最终分析验证研究将在三个地点进行：MSD，Johns 霍普金斯医院和儿童医院。多中心测试将包括测试熟练度样本， 20-日重现性研究和批间比较。此外，实时和加速稳定性测试 将在第四年进行。

This industrial-academic collaboration seeks to analytically validate a highly specific and sensitive multiplexed immunoassay for glial fibrillary acid protein (GFAP) and tau in serum and/or plasma to improve treatment decisions for neonates with hypoxic-ischemic encephalopathy (HIE). HIE, which affects approximately 1 in every 1,000 births, is a major cause of death and disability for children. Prompt whole body cooling of neonates improves outcomes in some cases; however, 24% of cooled neonates die from HIE, and 19% of survivors develop cerebral palsy despite best available treatment. Adjuvants to hypothermia promise to broaden the effectiveness of treatment. Clinical testing of these investigational adjuvants has been slow due to the difficulty in identifying neonates who are most likely to benefit from treatment. By prescreening neonates at the extremes of the injury spectrum, an objective simple blood test that stratifies neonates according to injury severity and monitors treatment responses would accelerate trials of investigational adjuvants. The multiplex employs a novel format with approximately 100-1,000x higher sensitivity than regular immunoassays, which improves quantitation in the clinically relevant ranges of these low abundance biomarkers. Our preliminary data demonstrate that our new assay format is sufficiently sensitive to measure GFAP and tau in serum and plasma. Collaborators at Johns Hopkins University (Drs. Everett and Northington) have demonstrated clinical utility of GFAP and tau as markers of injury severity and functional outcomes in HIE. This proposal seeks to fully develop and optimize the ultrasensitive multiplex using our phase-gated process, followed by extensive analytical validation. First-year tasks are extensive characterization of key critical reagents, assay fine-tuning, and establishment of analytical controls at key concentrations for stratifying neonates according to HIE severity. A risk analysis will be performed, and the effect of preanalytical factors and potential interferents will be assessed and mitigated. In year two, the design will be frozen and transferred to manufacturing. QC protocols for critical reagents will be established and executed, and components will be configured following MSD’s ISO 9001:2015-certified quality management system. Three kit lots at final design will be manufactured. Assay performance will be assessed throughout the project, and include testing of clinical samples. A formal analytical assay validation plan will be developed in collaboration with a clinical chemist (Dr. Sokoll from JHU), with input from the NIH Program Officers, and based on the 2018 FDA Bioanalytical Method Validation Guidance for Industry. The final analytical validation study will be performed at three sites: MSD, Johns Hopkins Hospital, and All Children’s Hospital. Multi-site testing will include testing of proficiency samples, a 20-day reproducibility study, and a lot-to lot comparison. Additionally, real-time and accelerated stability tests will be performed in year 4.