ULTRASENSITIVE CHEMILUMINESCENT ENZYME LINKED ASSAYS

描述：(改编自申请人的摘要) 建议的研究是开发超灵敏的化学发光分析 用免疫印迹和免疫印迹方法检测HIV-1抗体 囊性纤维化。申请人开发了一种专有的双酶- 结合多种作用的标记化学发光检测系统 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 一种新的HRP专有底物的化学发光。这个 申请人将用AP或HRP标记一对探针中的每个成员。 通过分析物在膜上结合两种探针，如在Western和 免疫印迹分析，将酶放在非常接近的位置， 促进化学发光增强。此外，申请人 将为这些检测开发一套化学发光试剂。这个 拟议中的研究应提供新一代化学发光 比比色法和比色法具有更高的灵敏度和特异度 其他化学发光测试。开发的技术将适用于 研究用于蛋白质和蛋白质的化学发光分析的试剂盒 核酸。

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.