Scientific Core 1:Human Clinical and Protein Engineering Core

科学核心1(SC1)。项目摘要/摘要 计划项目中的三个项目中的每个项目都需要经过提纯、原生和氧化修饰 脂蛋白，通过已知的临床随访获得人类临床材料，以及天然的、突变的或 在他们的研究中使用的特定位点的翻译后修饰的蛋白质。《人类临床》 材料与蛋白质工程核心(Science Core 1；SC1)将提供所有这些服务，允许 本计划每个项目中拟议研究的范围、规模和临床意义的扩大。 人类临床材料和蛋白质工程核心有三个主要目标：第一，核心将促进 通过提供以下服务，对项目研究人员进行人类临床和翻译研究：(I) 协调所有IRB文书工作、志愿者的广告/同意、患者与特定患者的联系 表型(例如，高Lp(A)水平)和采血服务。(Ii)使用标准化的SOP进行隔离和 下列人类生物材料的特性：全血(例如，用于分离PMN和MPO)； 血浆；血清；分离的脂蛋白(如极低密度脂蛋白、低密度脂蛋白、高密度脂蛋白和脂蛋白(A))。(三)单核细胞分离的配合 通过淘洗。(4)协调从以下地点获取人类临床标本档案(血浆、血清、DNA) 建议的临床研究的基因库队列。(V)可在以下地点获取人类临床标本 心血管诊断与预防中心。第二，核心将协助项目调查人员 通过提供以下服务进行蛋白质工程和抗体产生：(I)协助调查人员 设计、表达和纯化不同类别的野生型和突变型蛋白质。(Ii)发电 用于体内研究的低内毒素重组蛋白。(Iii)使用tRNA的正交氨基酸掺入 突变体以特异性地形成含有非天然氨基酸的单个重组蛋白(例如，硝基酪氨酸， 氯酪氨酸，N-ε-乙酰赖氨酸)。以及(4)协助产生多克隆和单克隆 抗体。最后，核心将作为方案调查人员的教育和培训中心，特别是 学生们和研究员们。将这些方面的工作和培训集中起来，将大大增强意义， 在本计划的每个项目中进行的研究的创新性和临床相关性。

Scientific Core 1(SC1). Project Summary/Abstract Each of the three Projects within the Program Project will require purified native and oxidatively modified lipoproteins, access to human clinical material with known clinical follow-up and a source of native, mutant or site-specific post-translationally modified proteins of interest to use in their studies. The Human Clinical Material & Protein Engineering Core (Scientific Core 1; SC1) will provide all of these services, allowing for expansion of the scope, scale and clinical significance of proposed studies in each Project of this Program. The Human Clinical Material & Protein Engineering Core has 3 primary objectives: First, the core will facilitate human clinical and translational studies of Program investigators by providing the following services: (i) Coordination of all IRB paperwork, advertising / consenting of volunteers, contact of patients with specific phenotypes (e.g. high LP(a) levels), and phlebotomy services. (ii) Using standardized SOP for isolation and characterization of the following human biological materials: whole blood (e.g. for PMN and MPO isolation); plasma; serum; isolated lipoproteins (e.g. VLDL, LDL, HDL and LP(a)). (iii) Coordination of monocyte isolation via elutriation. (iv) Coordination of access to archival human clinical specimens (plasma, serum, DNA) from the GeneBank cohort for proposed clinical studies. (v) Access to human clinical specimens available within the Center for Cardiovascular Diagnostics & Prevention. Second, the core will assist Program investigators with protein engineering and antibody generation by providing the following services: (i) assist investigators with design, expression, and purification of distinct classes of wild type and mutant forms of proteins. (ii) Generation of low endotoxin recombinant proteins for in vivo studies. (iii) Orthogonal amino acid incorporation using tRNA mutants to specifically form individual recombinant proteins harboring unnatural amino acids (e.g. nitrotyrosine, chlorotyrosine, Nε-acetyl-lysine). And (iv) assistance in generation of both polyclonal and monoclonal antibodies. Finally, the core will serve as an educational and training hub for Program investigators, especially students and fellows. Centralizing these aspects of the work and training will greatly enhance the significance, innovation, and clinical relevance of studies performed in each Project of this Program.