Point-of-care immunoassay panel for cardiotonic steroids

强心类固醇的即时免疫分析组合

• 批准号: 8779926
• 负责人: JAMES W LARRICK
• 金额: $ 22.5万
• 依托单位: PANORAMA RESEARCH, INC.
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2017-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8779926
• 关键词: Adverse effects; Affect; Affinity; Antibodies; Area; Binding; Biological Assay; Biological Markers; Blood; Blood Vessels; Body Fluids; Cardiac Glycosides; Clinical; Clinical Trials; Data; Detection; Development; Diagnosis; Diagnostic; Diagnostic tests; Digibind; Digoxin; Disease; Edema; Encephalopathies; Enzyme-Linked Immunosorbent Assay; Future; Goals; Health; Hypertension; Immune Sera; Immunoassay; K ATPase; Life; Literature; Liver Failure; Methods; Modeling; Monoclonal Antibodies; Morbidity - disease rate; Natriuresis; Outcome; Pathogenesis; Patients; Phase; Physiological; Placebos; Plasma; Pre-Eclampsia; Pregnancy; Pregnancy Complications; Proteinuria; Pump; Rattus; Recovery; Regulation; Relative (related person); Renal function; Research; Role; Sampling; Seizures; Sensitivity and Specificity; Signal Transduction; Specificity; Testing; Time; Translating; Urine; Woman; Work; analytical tool; antibody engineering; commercialization; digoxin-like factors; experience; fetal; inhibiting antibody; instrument; marinobufagenin; mortality; novel; point of care; pre-clinical; professor; response; tool; urinary

DESCRIPTION (provided by applicant): Preeclampsia (PE) is a serious complication of pregnancy manifested by high blood pressure, proteinuria, and edema, sometimes with encephalopathy, seizures, and hepatic failure. PE complicates from 5 to 10% of pregnancies, and is a major cause of maternal a fetal morbidity and mortality worldwide. Elevation in the circulating level of an endogenous "digoxin-like" factor (EDLF), an unknown substance that cross reacts with anti-digoxin antibodies and inhibits the NA+/K+ ATPase (NKA) was first noted in the 1980s. An extensive literature supports the hypothesis that increased levels of EDLF may be causative factor in the pathogenesis of hypertension. Recently, marinobufagenin (MBG), an endogenous cardiotonic steroid (CTS), has been identified as the EDLF. Plasma MBG is elevated over five-fold in pregnancies complicated by PE, suggesting a role pathophysiological role. Digibind, an anti-digoxin antiserum that cross-reacts MBG, provided significant in improvement in renal function relative to placebo among 51 patients with severe PE, with no adverse effects. Monoclonal antibodies to MBG are in clinical trials for PE. The lack of commercially available assays severely hampers research on the role MBG and at least two other cardiotonic steroids in PE and other diseases. The overall goal of this project is to develop an immunoassay panel for MBG and other potentially important endogenous cardiotonic steroids as a research tool and, ultimately, a clinical diagnostic for MBG levels in plasma and urine. In Phase I, we will identify monoclonal antibodies specific for each cardiotonic steroid demonstrating sensitivity and specificity. We will configure a competitive ELISA and will optimize it for the quantitation each CTS in plasma and urine. Finally, we will translate our competitive ELISA to a novel, automated immunoassay platform with ultrasensitive electrochemical detection. Phase II work will obtain data necessary for the submission of a 510k to support future commercialization.

描述（由申请人提供）：先兆子痫（PE）是一种严重的妊娠并发症，表现为高血压、蛋白尿和水肿，有时伴有脑病、癫痫发作和肝功能衰竭。PE使5%至10%的妊娠并发症，并且是全世界孕产妇和胎儿发病率和死亡率的主要原因。内源性“地高辛样”因子（EDLF）的循环水平升高是在20世纪80年代首次发现的，EDLF是一种与抗地高辛抗体交叉反应并抑制NA+/K+ ATP酶（NKA）的未知物质。大量文献支持EDLF水平升高可能是高血压发病机制中的致病因素的假设。近年来，内源性强心类固醇（CTS）--海蟾毒配基（MBG）已被鉴定为EDLF。血浆MBG在妊娠合并PE中升高超过5倍，表明其具有病理生理作用。Digibind是一种与MBG交叉反应的抗地高辛抗血清，在51例重度PE患者中，与安慰剂相比，Digibind显著改善了肾功能，无不良反应。针对MBG的单克隆抗体正在进行PE的临床试验。缺乏商业上可获得的测定严重阻碍了对MBG和至少两种其他强心类固醇在PE和其他疾病中的作用的研究。该项目的总体目标是开发 MBG和其他潜在重要的内源性强心类固醇的免疫测定面板作为研究工具，并最终用于血浆和尿液中MBG水平的临床诊断。在第一阶段，我们将确定单克隆抗体的每一个强心类固醇证明敏感性和特异性。我们将配置竞争性ELISA，并将优化其用于定量血浆和尿液中的每种CTS。最后，我们将把我们的竞争性ELISA转化为一种新型的、具有超灵敏电化学检测的自动化免疫分析平台。第二阶段的工作将获得提交510 k所需的数据，以支持未来的商业化。