CHEMILUMINESCENT WESTERN BLOT SYSTEM FOR AIDS DIAGNOSIS

用于艾滋病诊断的化学发光蛋白质印迹系统

• 批准号: 3489330
• 负责人: JOHN C VOYTA
• 金额: $ 5万
• 依托单位: TROPIX, INC.
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-09-30 至 1991-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3489330
• 关键词: AIDS; AIDS /HIV diagnosis; diagnosis design /evaluation; luminescence; western blottings

The goal of Phase I of this proposal is to establish a chemiluminescent detection system for AIDS Western blotting based on the chemiluminescent substrate for alkaline phosphatase, AMPPD (trademark). One advantage of this system will be increased sensitivity, over existing Western blots tests, which result in stained membrane strips. Also, a permanent record of results will be produced directly on film. A model Western blot system using bovine serum albumin (BSA) and commercially available antibodies will be used to develop a new luminescence amplifying system which will be necessary for nitrocellulose membrane. Blocking conditions and sensitivity will also be optimized. The luminescent amplifying system will result in a chemiluminescent emission with a maximum at approximately 540 nm. This wavelength is a good match for capturing the chemiluminescent signal on Polaroid instant black and white film. The optimal conditions developed with the BSA model system will then be applied to components of a DuPont HIV Western blot kit, using the kit's noninfectious controls to test the performance and sensitivity of the chemiluminescent system. The results obtained on instant film will be suitable for

该提案第一阶段的目标是建立一个发光 基于荧光免疫印迹的艾滋病检测系统 碱性磷酸酶底物AMPPD（商标）。的一个优点 该系统将比现有的蛋白质印迹法更灵敏， 测试，导致染色的膜条。此外，永久记录 结果将直接在胶片上产生。一个模型Western印迹系统 使用牛血清白蛋白（BSA）和市售抗体 用于开发一种新的发光放大系统， 硝酸纤维素膜所必需的。阻塞条件和灵敏度 也将得到优化。发光放大系统将导致 在约540 nm处具有最大值的荧光发射。这 波长是用于捕获光致发光信号的良好匹配。 宝丽来即时黑色和白色电影。开发的最佳条件 与BSA模型系统，然后将应用于组件的杜邦 HIV蛋白质印迹试剂盒，使用试剂盒的非感染性对照品检测 的性能和灵敏度。结果 在即时电影上获得的将适合于