Mass Spectrometry in Clinical Diagnosis of Nerve Agent Exposure

质谱法在神经毒剂暴露临床诊断中的应用

基本信息

批准号：
7470621
负责人：
OKSANA LOCKRIDGE
金额：
$ 45.54万
依托单位：
UNIVERSITY OF NEBRASKA MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-09-27 至 2010-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7470621
关键词：
Active Sites Adopted Albumins Binding Biological Assay Biological Markers Blood Blood Tests Blood specimen Butyrylcholinesterase Characteristics Charge Cholinesterases Clinical Detection Diagnosis Diagnostic Diagnostic Procedure Digestion Dissociation Dose Emergency Situation Endopeptidases Exposure to Funding Opportunities Goals Hospitals Human In Vitro Ions Label Life Liquid Chromatography Mass Spectrum Analysis Measurement Measures Medical Methods Monitor Mus Parents Pattern Pepsin A Peptide Hydrolases Peptides Pesticides Plasma Proteins Reaction Research Research Personnel Research Project Grants Sampling Sarin Serine Soman Suicide attempt Symptoms Testing Toxicant exposure Trypsin Tyrosine United States adduct clinical Diagnosis covalent bond enzyme activity human subject inhibitor/antagonist mass spectrometer nerve agent novel diagnostics pesticide exposure programs rapid diagnosis response tandem mass spectrometry toxicant

项目摘要

DESCRIPTION (provided by applicant): The goal of this project is to develop a rapid and sensitive assay for nerve agent exposure in humans. Existing assays do not detect low dose exposure and cannot reassure the public that no exposure has occurred. Two proteins in blood, butyrylcholinesterase and albumin, will be tested as biomarkers of nerve agent exposure in our new assays. Butyrylcholinesterase is an established biomarker in assays that measure loss of enzyme activity. However, measurement of butyrylcholinesterase activity does not identify the inhibitor as a nerve agent, and does not reveal low dose exposure. Albumin is a new biomarker whose utility as a biomarker has not previously been tested in humans, though it shows promise in mice. Our proposed new assays use MALDI-TOF and tandem mass spectrometry to detect peptides covalently attached to nerve agent. The Selective Reaction Monitoring method is expected to detect as little as 100 femtomoles of covalently bound nerve agent in human plasma. The assays will be developed with human plasma treated with soman and sarin in vitro. The utility of the assays will be evaluated by testing plasma from human subjects exposed to nerve agent simulants. The specific aims are 1) Develop a method to rapidly diagnose exposure to nerve agents, by treating human blood with soman and sarin, and analyzing samples in the mass spectrometer. The method will look for peptides of albumin-nerve agent adducts and of butyrylcholinesterase-nerve agent adducts. 2) Develop a method to rapidly diagnose exposure to nerve agent simulants, by testing blood from humans accidentally exposed to organophosphorus pesticides, or from attempted suicide cases who ingested organophosphorus pesticides. Mass spectrometry will be used to identify exposure. This specific aim differs from aim #1 in that humans will have been exposed to nerve agent simulants; nerve agent simulants must be studied because humans exposed to nerve agents are not available. The new diagnostic methods have the potential for saving lives in an emergency because correct, rapid diagnosis indicates the appropriate medical treatment.

描述（由申请人提供）：该项目的目标是开发一种快速、灵敏的人体神经毒剂暴露检测方法。现有的检测方法无法检测低剂量暴露，也无法向公众保证没有发生暴露。血液中的两种蛋白质，丁酰胆碱酯酶和白蛋白，将在我们的新测定中作为神经毒剂暴露的生物标志物进行测试。丁酰胆碱酯酶是测量酶活性损失的测定中的既定生物标志物。然而，丁酰胆碱酯酶活性的测量并不能将抑制剂识别为神经毒剂，也不能揭示低剂量暴露。白蛋白是一种新的生物标志物，其作为生物标志物的效用此前尚未在人体中进行过测试，尽管它在小鼠身上显示出前景。我们提出的新检测方法使用 MALDI-TOF 和串联质谱法来检测共价连接到神经毒剂的肽。选择性反应监测方法预计可检测人血浆中低至 100 飞摩尔的共价结合神经毒剂。这些检测方法将在体外用梭曼和沙林处理过的人血浆进行开发。将通过测试暴露于神经毒剂模拟物的人类受试者的血浆来评估这些测定的实用性。具体目标是 1) 通过用梭曼和沙林处理人体血液并在质谱仪中分析样本，开发一种快速诊断神经毒剂暴露的方法。该方法将寻找白蛋白-神经剂加合物和丁酰胆碱酯酶-神经剂加合物的肽。 2) 开发一种快速诊断神经毒剂模拟物暴露的方法，通过测试意外接触有机磷农药的人类或摄入有机磷农药的自杀未遂病例的血液。质谱分析法将用于识别暴露情况。这一具体目标与目标#1 的不同之处在于，人类将接触神经毒剂模拟物；必须研究神经毒剂模拟物，因为无法接触到神经毒剂的人类。新的诊断方法有可能在紧急情况下挽救生命，因为正确、快速的诊断表明了适当的医疗治疗。

项目成果

期刊论文数量（2）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Production of ES1 plasma carboxylesterase knockout mice for toxicity studies.

生产 ES1 血浆羧酸酯酶敲除小鼠用于毒性研究。

DOI：
发表时间：
2011-11-21
期刊：
影响因子：
4.1
作者：
Duysen, Ellen G;Koentgen, Frank;Williams, Gareth R;Timperley, Christopher M;Schopfer, Lawrence M;Cerasoli, Douglas M;Lockridge, Oksana
通讯作者：
Lockridge, Oksana

Aging pathways for organophosphate-inhibited human butyrylcholinesterase, including novel pathways for isomalathion, resolved by mass spectrometry.

通过质谱法解析了有机磷抑制的人丁酰胆碱酯酶的衰老途径，包括异马拉硫磷的新途径。