CORE--LIPID AND LIPOPROTEIN BIOCHEMISTRY

核心--脂质和脂蛋白生物化学

• 批准号: 6302159
• 负责人: JOHN L VANDEBERG
• 金额: $ 42.63万
• 依托单位: TEXAS BIOMEDICAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-01-01 至 2000-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6302159
• 关键词: apolipoproteins; biomedical facility; blood chemistry; blood lipid; blood lipoprotein; blood preservation; cholesterol; chylomicrons; densitometry; density gradient ultracentrifugation; dietary lipid; electron microscopy; enzyme linked immunosorbent assay; gel electrophoresis; high density lipoproteins; high performance liquid chromatography; low density lipoprotein; nutrition related tag; phenotype; triglycerides; very low density lipoprotein; western blottings

The Lipid and Lipoprotein Biochemistry Core Laboratory will serve all projects on a highly interactive and collaborative basis. The primary role of this core unit is to provide phenotypic data on measures of plasma lipoproteins. This core unit also is responsible for processing and storing the blood samples used by the core unit and by Project 1. Quantitative measures of lipoprotein phenotypes will be made by clinical chemical and immunochemical techniques. Standard clinical chemical procedures will be used to quantify serum TC and HDL-C after precipitation. Immunoturbidimetric and other immunochemical techniques will be used to measure serum concentrations of apoAI, apoB, apoE, LP(A) particles bearing specific isoforms. In addition, we will determine apo(a) isoform phenotypes. The gradiant gel electrophoresis (GGE) technique, established in our laboratory for the specific purpose of genetic research involving large numbers of samples, will enable estimates of quantities of cholesterol in each of four size-resolved HDL particle classes: HDL1a, HDL1b HDL2, and HDL3. In addition, the GGE technique will be used to determine the distribution of cholesterol among four size-resolved LDL particle classes, as well as the median particle diameter and diameter of the predominant LDL species in each sample. Blood processing will involve isolating serum for phenotypic measures, processing it for storage as 60 mul aliquots, and storing clots as a backup source of DNA. Sera and clots will be catalogued in the computerized inventory database, maintained at -80 degrees Centigrade, and retrieved as needed. Blood from 750 patients and inbred progeny projected to undergo the dual dietary challenge will be processed and assayed for lipoprotein phenotypes on the occasions of the three dietary samplings. In addition, LP(a) phenotypes, apo(a) phenotypes, and LDL cholesterol distributions will be assayed on 1,800 samples from non-inbred animals that underwent the dual dietary challenge during the current project period.

脂质和脂蛋白生物化学核心实验室将为所有人服务 项目在高度互动和协作的基础上进行。主要角色 该核心单元的目的是提供血浆测量的表型数据 脂蛋白。该核心单元还负责处理和 存储核心单元和项目 1 使用的血液样本。 脂蛋白表型的定量测量将通过 临床化学和免疫化学技术。标准临床 化学程序将用于量化血清 TC 和 HDL-C 后 沉淀。免疫比浊法和其他免疫化学技术 将用于测量apoAI、apoB、apoE、LP(A)的血清浓度 具有特定异构体的颗粒。此外，我们将确定apo(a) 同工型表型。 我们建立的梯度凝胶电泳 (GGE) 技术 用于涉及大型基因研究的特定目的的实验室 样本数量，将能够估计体内胆固醇的数量 四个尺寸分辨 HDL 颗粒类别中的每一个：HDL1a、HDL1b HDL2 和 高密度脂蛋白3。此外，GGE技术将用于确定 四种尺寸分辨的 LDL 颗粒类别中胆固醇的分布， 以及中值​​粒径和主要颗粒的直径 每个样本中的 LDL 种类。 血液处理将涉及分离血清以检测表型 测量，将其处理为 60 μl 等分试样储存，并储存凝块 作为 DNA 的备份来源。血清和凝块将编目于 计算机化库存数据库，保持在-80摄氏度，以及 根据需要检索。 来自 750 名患者和近交后代的血液预计将接受 将处理双重饮食挑战并检测脂蛋白 三个饮食样本的表型。此外， LP(a) 表型、apo(a) 表型和 LDL 胆固醇分布 将对来自非近交动物的 1,800 个样本进行分析 当前项目期间的双重饮食挑战。