Demonstration of a homogeneous noncompetitive immunoassay based on bioluminescence resonance energy transfer
Demonstration of a homogeneous noncompetitive immunoassay based on bioluminescence resonance energy transfer
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DOI：
10.1006/abio.2000.4924
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发表时间：
2001-02-01
影响因子：
2.9
通讯作者：
Nagamune, T
中科院分区：
文献类型：
--
作者：
Arai, R;Nakagawa, H;Nagamune, T
我们描述了一种基于抗体可变区抗原依赖重结合的非竞争性均相生物发光免疫分析方法(开放夹心生物发光免疫分析，OS-BLIA)，通过生物发光共振能量转移(BRET)监测了抗体重链片段(V-H)-肾形荧光素酶(Rluc)和抗体轻链片段(V-L)增强型黄色荧光蛋白(EYFP)两个嵌合蛋白的重新结合。将试剂与样品简单混合后，观察到BRET的抗原依赖性增加，其可测量的浓度范围为0.1，类似于10µg/ml的抗原鸡蛋溶菌酶，与我们基于荧光共振能量转移(FRET)的可比分析方法相比，灵敏度提高了10倍，可能是由于试剂浓度的降低，(C)2001学术出版社。
We describe a noncompetitive homogeneous bioluminescent immunoassay based on the antigen-dependent reassociation of antibody variable domains (open sandwich bioluminescent immunoassay, OS-BLIA), The reassociation of two chimeric proteins, an antibody heavy-chain fragment (V-H)-Renilla luciferase (Rluc) and an antibody light-chain fragment (V-L)-enhanced yellow fluorescent protein (EYFP), was monitored by a bioluminescence resonance energy transfer (BRET) between the two. Upon simple mixing of the reagents with the sample, an antigen-dependent increase in BRET was observed with a measurable concentration range of 0.1 similar to 10 mug/ml antigen hen egg lysozyme, Compared with our comparable assays based on fluorescence resonance energy transfer (FRET), a 10-fold improvement in the sensitivity was attained, probably due to a reduction in reagent concentration, (C) 2001 Academic Press.