Development of a homegeneous immunodiagnostics system utilizing luminescence wavelength transformation by energy transfer

为了建立灵敏的均相免疫分析法，尝试验证利用发光能量传递和发光波长变换的均相免疫分析法原理。将不同量的荧光标记抗体与嵌合蛋白A-Vargula荧光素酶和底物荧光素混合后，荧光光谱出现明显红移，可能是由于有效的发光能量转移。也可以用带长通滤光器的微滴孔式光度计检测。标记抗体的数量与红移波长发光的程度有良好的相关性，表明了新型发光均相免疫分析法的可行性。为了获得更好的灵敏度，我们制备了一种新的G-Vargula荧光素酶嵌合蛋白，其灵敏度是a -荧光素酶的2倍。因此，该结果暗示了采用表位标记荧光素酶和荧光标记抗体的新抗原检测方法的可能性。

To develop a sensitive homogeneous immunoassay, verification of a homogeneous immunoassay principle which utilize luminescence energy transfer and transformation of luminescence wavelength, was attempted.When variable amount of fluorescent labeled antibody was mixed with chimeric protein A-Vargula luciferase and substrate luciferin, marked red shift in luminescence spectre possibly due to efficient luminescence energy transfer was detected. It was also detectable with a microtiter well-type luminometer with long pass filter unit. The amount of labeled antibody correlated well with the extent of luminescence with red-shifted wavelength, which suggests the feasibility of new luminescent homogeneous immunoassay. To obtain better sensitivity in the assay, a novel chimeric protein of protein G-Vargula luciferase was made and shown to have 2-fold sensitivity than protein A-luciferase. The result thus implies a possibility of new antigen detection method employing epitope-tagged luciferase and fluorescent-labeled antibody.