ULTRASENSITIVE CHEMILUMINESCENT ENZYME LINKED ASSAYS

超灵敏化学发光酶联检测

• 批准号: 2147544
• 负责人: Hashem Akhavan-Tafti
• 金额: $ 38.25万
• 依托单位: LUMIGEN, INC.
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-03-01 至 1997-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2147544
• 关键词: AIDS /HIV test; alkaline phosphatase; antiviral antibody; avidin; biotin; chloride channels; cystic fibrosis; fluoresceins; gel electrophoresis; haptens; horseradish peroxidase; human immunodeficiency virus 1; immunoconjugates; luminescence; method development; nucleic acid probes; oligonucleotides; point mutation; southern blotting; western blottings

DESCRIPTION: (Adapted from the applicant's abstract) The objective of the proposed research is to develop ultrasensitive chemiluminescent assays for HIV-1 antibodies by Western blotting and immunoblotting assays for cystic fibrosis. The applicant has developed a proprietary dual-enzyme- label chemiluminescent detection system combining the actions of alkaline phosphatase (AP) and horse radish peroxidase (HRP). AP hydrolyzes a substrate to produce a luminescence enhancer for generating chemiluminescence from a new proprietary substrate for HRP. The applicant will label each member of a pair of probes with AP or HRP. Binding of both probes by an analyte on a membrane, as in Western and immunoblotting assays, places the enzymes in close proximity, facilitating chemiluminescence enhancement. In addition, the applicant will develop sets of chemiluminescent reagents for these assays. The proposed research should provide a new generation of chemiluminescent tests with improved sensitivity and specificity over colorimetric and other chemiluminescent tests. Technology developed will be applicable to research kits for performing chemiluminescent assays for proteins and nucleic acids.

描述：（改编自申请人的摘要） 拟议的研究是开发超灵敏的荧光分析 通过Western印迹和免疫印迹分析检测HIV-1抗体， 囊性纤维化申请人开发了一种专有的双酶- 标签发光检测系统，其结合以下动作： 碱性磷酸酶（AP）和辣根过氧化物酶（HRP）。AP 水解底物以产生发光增强剂， 化学发光从一个新的专有底物HRP。的 申请人将用AP或HRP标记一对探针的每个成员。 两种探针通过膜上的分析物的结合，如在Western和Western中。 免疫印迹分析，将酶放置在非常接近的位置， 促进化学发光增强。此外，申请人 将为这些检测开发一套荧光试剂。的 拟议的研究将提供新一代的荧光 与比色法相比具有改进的灵敏度和特异性的测试， 其他荧光测试。开发的技术将适用于 涉及用于对蛋白质进行荧光测定的研究试剂盒， 核酸