Protein and PCR Mass Spectrometry

PSWRCE蛋白和PCR MS核心将在以下项目上合作： 特定目标i -伯克霍尔德菌、球孢子菌和储库疫苗项目的蛋白质组学分析 识别用作诊断或疫苗候选物的蛋白质生物标志物（来自血清、尿液和肺） 匀浆，必要时消耗大量小鼠/人蛋白质） 对预分级样品进行比较蛋白质组学（例如，胞外蛋白或细胞壁 生物体的两种不同菌株的蛋白质，或野生型与突变型） 构建“血粉”蛋白质文库用于鉴定蜱虫中的宿主脊椎动物蛋白质 具体目标2 -开发诊断分析 使用Sp. Aim i的生物标志物ID开发用于病原体鉴定的生物亲和MS 使用MS指导靶向ELISA或其他荧光测定法的开发（表征表达的 在整个试验开发过程中通过MS测定蛋白抗原、抗体和标记抗体） 比较抗体与小配体在生物亲和MS中捕获抗原的情况 3 - Ibis TSOOO PCR-MS生物传感器应用于储库疫苗、伯克霍尔德菌和球孢子菌 ·PCR MS（在线粒体DNA上）用于载体所喂养的脊椎动物宿主的ID（例如，蜱若虫 以黑尾鹿为食）和蜱传病原体ID <$PCR MS用于病原体的快速ID和菌株分型，以及毒素存在的确认 基因，抗生素抗性基因，选择性毒力因子基因 具体目标4 -蛋白质复合物的表征（未补偿协同活性） 结构表征大蛋白质-蛋白质复合物（例如，参与毒力的皮利，组装 的噬菌体尾纤维蛋白用于生物传感器），使用一种改良的QTOF MS 抗原的发现与诊断测定开发、PCR-MS、生物亲和MS和 大蛋白质-蛋白质复合物的结构表征是核心的独特能力。

The PSWRCE Protein and PCR MS Core will collaborate on the following projects: Specific Aim i - Proteomics Analyses for Burkholderia, Coccidioides, and Reservoir Vaccines Projects ¿ Identify protein biomarkers for use as diagnostic or vaccine candidates (from serum, urine, and lung homogenates, with depletion of abundant mouse/human proteins as necessary) ¿ Perform comparative proteomics on pre-fractionated samples (e.g., extracellular proteins or cell wall proteins of two different strains of an organism, or wild type vs. mutant) ¿ Build "blood meal" protein library to use to identify host vertebrate proteins in tick nymphs Specific Aim 2 - Development of Diagnostic Assays ¿ Use biomarker IDs of Sp. Aim i to develop bioaffinity MS for pathogen identification ¿ Use MS to guide development of targeted ELISA or other fluorescence assays (characterize expressed protein antigens, antibodies, and labeled antibodies by MS throughout development of assay) ¿ Compare antibodies vs. small ligands for capture of antigens in bioaffinity MS Specific Aim 3 - Ibis TSOOO PCR-MS Biosensor Applied to Reservoir Vaccines, Burkholderia & Coccidioides ¿ PCR MS (on mitochondrial DNA) for ID of vertebrate host on which vector has fed (e.g., tick nymph fed on mule deer) and ID of tick-borne pathogenic organism present ¿ PCR MS for rapid ID and strain typing of pathogens, as well as confirmation of the presence of toxin genes, antibiotic resistance genes, selected virulence factor genes Specific Aim 4 - Characterization of Protein Complexes (uncompensated synergistic activity) ¿ Structurally characterize large protein-protein complexes (e.g., pili involved in virulence, assemblies of bacteriophage tail fiber proteins useful for biosensors) using a one-of-a-kind modified QTOF MS The antigen discovery coupled to support of diagnostic assay development, PCR-MS, bioaffinity MS, and structural characterization of large protein-protein complexes are unique capabilities of the core.