AB Sciex Q-Trap 5500 LC/MS/MS

描述（由申请人提供）：该提案要求为Eksigent的Applied Biosystems SCIEX QTRAP 5500质谱仪提供资金，该质谱仪配有Ultra 2D纳米液相色谱（LC）系统，该系统专门用于临床/转化研究，其中使用称为多反应监测（MRM）的方法获得单个或多个蛋白质的绝对定量。MRM测定产生与ELISA相同的质量数据，但不需要抗体。因此，MRM缩短了临床样品组中一种或多种蛋白质的定量测定开发的时间和费用。此外，多重MRM测定消除了对抗体阵列的需要，抗体阵列在技术上具有挑战性并且通常是有限的。我们将使用QTRAP 5500 LC/MS/MS专门用于大型临床队列中的MRM测定，其中该设备的独特方面至关重要。例如，（i）大量蛋白质（40+）及其修饰形式同时测定，（ii）广泛的蛋白质组覆盖（1 e7+）需要最大的灵敏度，（iii）必须检查大样本集（500+），（iv）只有非常少量的不可替代的样本可用。一组非常有经验的研究人员将使用QTRAP 5500进行针对性研究，以解决关键的临床问题。七个团队构成了主要的用户，包括来自Jennifer货车Eyk指导的约翰霍普金斯CTSA/ITCR生物标志物开发中心的研究人员和MS专家，以及Anne Murphy、艾伦埃弗雷特和James Casella、Josef Coresh、M.克莉丝汀津克和大卫格雷厄姆，哈里（哈尔）迪茨以及杰拉尔德哈特。研究人员将共同努力，最大限度地利用仪器，并确保高质量和有效的数据分析。这些项目涉及心脏病学、血管生物学、中风、糖尿病、镰状细胞性贫血、囊性纤维化、癌症和艾滋病毒等临床问题。每个项目都将对临床医学产生影响。 公共卫生相关性：该提案要求为能够定量临床样本中特定蛋白质的质谱仪提供资金。这种特殊仪器的独特之处在于：（i）能够同时分析大量蛋白质及其修饰形式 (40+），（ii）增加的灵敏度，使蛋白质组覆盖率最大化（1 e7+），（iii）能够分析大量样品（500+）和（iv）非常少量的人类临床样品。这些项目解决了心脏病学、血管生物学、中风、糖尿病、镰状细胞性贫血、囊性纤维化、癌症和艾滋病毒等重要而紧迫的临床问题。每个项目都将对临床医学产生影响。

DESCRIPTION (provided by applicant): This proposal requests funds for an Applied Biosystems SCIEX QTRAP 5500 mass spectrometer with an Ultra 2D nano liquid chromatographic (LC) system from Eksigent, This system has been chosen specifically for the clinical/translational studies in which absolute quantification of a single or many proteins is obtained using an approach known as multiple reaction monitoring (MRM). MRM assays produce the same quality data as a ELISA but without the need for antibodies. Thus, MRM condenses the time and expense of quantitative assay development for one or more protein in clinical sample sets. Furthermore, multiplex MRM assays eliminate the need for antibody arrays, which are technically challenging and often limited. We will be using the QTRAP 5500 LC/MS/MS specifically for MRM assays in the context of large clinical cohorts where the unique aspects of this equipment are essential. Examples include situations where (i) a large number of proteins (40+), and their modified forms are assayed simultaneously, (ii) maximal sensitivity is needed for broad proteome coverage (1e7+), (iii) large sample sets (500+) must be examined and (iv) only very small quantities of irreplaceable samples are available. A group of very experienced investigators with targeted studies to address key clinical questions will use the QTRAP 5500. Seven teams constitute the major users and include researchers and MS experts from the Johns Hopkins CTSA/ITCR Biomarker Development Center directed by Jennifer Van Eyk, and the laboratories of Anne Murphy, Allen Everett and James Casella, Josef Coresh, M. Christine Zink and David Graham, Harry (Hal) Dietz as well as Gerald Hart. The researchers will work collaboratively to maximize the instrument use and to ensure high quality and effective data analysis. The projects address clinical questions in cardiology, vascular biology, stroke, diabetes, sickle cell anemia, cystic fibrosis, cancer, and HIV. Each project is poised to impact clinical medicine. PUBLIC HEALTH RELEVANCE: This proposal requests funds for a mass spectrometer capable of quantifying specific proteins in clinical samples. The unique aspects of this particular instrument is it's (i) ability to simultaneously analyze a large number of proteins and their modified forms (40+), (ii) increased sensitivity which maximizes proteome coverage (1e7+), (iii) capability to analyze large sample numbers (500+) with (iv) very small quantities of human clinical samples. The projects address important and pressing clinical questions in cardiology, vascular biology, stroke, diabetes, sickle cell anemia, cystic fibrosis, cancer, and HIV. Each project is poised to impact clinical medicine.