AB Sciex Q-Trap 5500 LC/MS/MS

描述（由申请人提供）：本提案要求为应用生物系统SCIEX QTRAP 5500质谱计提供资金，该质谱计配备Eksigent的超2D纳米液相色谱（LC）系统，该系统专门用于临床/转化研究，其中使用称为多重反应监测（MRM）的方法获得单个或多个蛋白质的绝对定量。MRM分析产生与ELISA相同质量的数据，但不需要抗体。因此，MRM压缩了临床样品组中一种或多种蛋白质定量分析开发的时间和费用。此外，多重MRM分析消除了对抗体阵列的需求，抗体阵列在技术上具有挑战性，而且通常是有限的。我们将在大型临床队列中使用QTRAP 5500 LC/MS/MS专门用于MRM分析，其中该设备的独特方面是必不可少的。例子包括：(i)同时分析大量蛋白质（40+）及其修饰形式，（ii）广泛的蛋白质组覆盖需要最大的灵敏度（1e7+）， （iii）必须检查大量样品集（500+），以及（iv）只有非常少量的不可替代的样品可用。一组非常有经验的研究人员将使用QTRAP 5500进行有针对性的研究，以解决关键的临床问题。七个团队构成了主要用户，包括来自约翰霍普金斯CTSA/ITCR生物标志物开发中心的研究人员和MS专家，由Jennifer Van Eyk领导，以及Anne Murphy， Allen Everett和James Casella， Josef Coresh， M. Christine Zink和David Graham， Harry (Hal) Dietz以及Gerald Hart的实验室。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、癌症和艾滋病毒的临床问题。每个项目都准备影响临床医学。

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.