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固相时间分辨荧光免疫标记技术研究
Study of Solid-Phase Time-Resolved Fluorescence Label Immunoassay
【摘要】 通过固相时间分辨荧光免疫分析双功能螯合剂 4 ,7 二氯磺基苯 1,10菲罗啉 2 ,9 二羧酸标记抗 乙型肝炎表面抗体 (HBsAb)IgG实验 ,对于BCPDA标记蛋白质的方法进行了研究。结果表明 :BCPDA在相对温和条件下能与蛋白质反应 ,反应后蛋白质的相对生物活性高于 78% ,标记比为 2 3~ 5 5 ,蛋白回收率达6 0 %以上。在一定条件下与铕离子形成稳定的BCPDA Eu3 + (HBsAb)IgG标记物。利用自建的分析方法 ,测定了标记过程的有关参数。并对标记物的某些光学特性进行了研究。
【Abstract】 This paper describes optimal conditions for HBsAbIgG labeling with a new fluorescence probe, 4,7-bis-chorosulfophenyl-1,10-phenanthroline-2,9-dicarboxylic acid(BCPDA)for the solid phase time-resolved fluorimmunoassay (TRFIA). The result of experiment under states clearly that BCPDA may react with protein under relative mild condition. The relative bioactivity of reacted protein was more than 80%. The labeling molar ratio of BCPDA for HBsAbIgG was 45-70. The recovery was higher than 80%. Protein-BCPDA-Eu 3+ complex is stable. It can emit very high fluorescence intensity with very long fluorescence life times. The fluorescence of Protein-BCPDA-Eu 3+ complex has a very large stokes shift (270 nm). The emission band at 611.2 nm is very narrow. The research provides the base for developing non-isotopic immunoassay technique and clinical medical diagnosis.
【Key words】 Solid-phase time-resolved fluorimmunoassay; Europium labels; Anti-hepatits B surface; 4,7-bis-chorosulfophenyl-1,10-phenanthroline-2,9-dicarboxylic acid;
- 【文献出处】 光谱学与光谱分析 ,Spectroscopy and Spectral Analysis , 编辑部邮箱 ,2004年12期
- 【分类号】R392.3
- 【被引频次】22
- 【下载频次】388