Scientific Core 1:Human Clinical and Protein Engineering Core

科学核心1：人类临床和蛋白质工程核心

• 批准号: 9049529
• 负责人: Joseph DiDonato
• 金额: $ 14.96万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9049529
• 关键词: 3-nitrotyrosine; Advertising; Amino Acids; Animal Disease Models; Antibodies; Antigens; Aorta; Biochemical; Biocompatible Materials; Biological Assay; Blood Vessels; Cardiovascular Diseases; Cardiovascular system; Caring; Carotid Arteries; Clinical; Clinical Engineering; Clinical Research; Consent; DNA; Diagnostic; Education; Endotoxins; Funding; Generations; Goals; High Density Lipoproteins; Human; In Vitro; Individual; Inflammation; Institutional Review Boards; Lipoprotein (a); Lipoproteins; Low-Density Lipoproteins; Lysine; Molecular; Molecular Genetics; Monoclonal Antibodies; Myocardial Infarction; Names; Patients; Peripheral; Phenotype; Plasma; Prevention; Protein Engineering; Proteins; Recombinant Proteins; Registries; Research; Research Personnel; Resources; Sampling; Serum; Services; Site; Source; Specimen; Structure; Students; Techniques; Tissues; Training; Training Programs; Training and Education; Transfer RNA; Venous blood sampling; Very low density lipoprotein; Whole Blood; Work; abstracting; clinical material; clinically relevant; clinically significant; cohort; design; follow-up; human biological material; in vivo; innovation; interest; liver biopsy; lung injury; monocyte; mutant; myocardial biopsy; oxidation; polyclonal antibody; programs; protein expression; repository; translational study; unnatural amino acids; volunteer

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.

科学核心 1（SC1）。项目概要/摘要 计划项目中的三个项目中的每一个都需要纯化的天然和氧化修饰的 脂蛋白，获得具有已知临床随访的人类临床材料以及天然、突变或 在他们的研究中使用感兴趣的位点特异性翻译后修饰蛋白质。人类临床 材料与蛋白质工程核心（科学核心 1；SC1）将提供所有这些服务，允许 扩大本计划每个项目中拟议研究的范围、规模和临床意义。 人类临床材料和蛋白质工程核心有 3 个主要目标：首先，该核心将促进 项目研究者通过提供以下服务进行人类临床和转化研究：(i) 协调所有 IRB 文件、广告/志愿者同意、与特定患者的联系 表型（例如高 LP(a) 水平）和放血服务。 (ii) 使用标准化 SOP 进行隔离和 以下人类生物材料的表征：全血（例如用于 PMN 和 MPO 分离）； 等离子体;血清;分离脂蛋白（例如 VLDL、LDL、HDL 和 LP(a)）。 (iii) 单核细胞分离的协调 通过淘析。 (iv) 协调从机构获取人类临床档案标本（血浆、血清、DNA） 用于拟议临床研究的 GeneBank 队列。 (v) 获取可用的人类临床样本 心血管诊断与预防中心。其次，核心将协助计划调查人员 通过提供以下服务来进行蛋白质工程和抗体生成：(i) 协助研究人员 不同类别的野生型和突变型蛋白质的设计、表达和纯化。 (二) 世代 用于体内研究的低内毒素重组蛋白。 (iii) 使用 tRNA 进行正交氨基酸掺入 突变体专门形成含有非天然氨基酸（例如硝基酪氨酸， 氯酪氨酸、Nε-乙酰基-赖氨酸）。 (iv) 协助产生多克隆和单克隆 抗体。最后，该核心将作为项目研究人员的教育和培训中心，特别是 学生和研究员。集中这些方面的工作和培训将大大增强意义， 本计划每个项目中进行的研究的创新性和临床相关性。