| 固相时间分辨荧光免疫标记技术研究 |
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| 引用本文： | 潘利华,周誓红,孙文伟,谢文兵,赵超.固相时间分辨荧光免疫标记技术研究[J].光谱学与光谱分析,2004,24(12):1601-1604. |
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| 作者姓名： | 潘利华 周誓红 孙文伟 谢文兵 赵超 |
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| 作者单位： | 中国科学院长春应用化学研究所,国家电化学光谱分析研究中心,中国科学院稀土化学和物理开放实验室,吉林,长春,130022;吉林大学中日联谊医院,吉林,长春,130031 |
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| 基金项目： | 国家自然科学基金 (30 0 70 71 4 )资助项目 |
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| 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23～55，蛋白回收率达60％以上。在一定条件下与铕离子形成稳定的BCPDA-Eu^3 (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。
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| 关 键 词： | 固相时间分辨荧光免疫 铕标记 抗-乙型肝炎表面抗体 4 7-二氯磺基苯-1 10菲罗啉-2 9-二羧酸 |
| 文章编号： | 1000-0593(2004)12-1601-04 |
| 修稿时间： | 2003年2月8日 |
Study of Solid-Phase Time-Resolved Fluorescence Label Immunoassay |
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| PAN Li-hua ,ZHOU Shi-hong ,SUN Wen-wei ,XIE Wen-bing ,ZHAO Chao . National Analytical Research Center of Electrochemistry and Spectroscopy,Key Laboratory of Rare Earth Chemistry and Physics,Changchun Institute of Applied Chemistry,Chinese Academy of Sciences,Changchun ,China . China-Japan Union Hospital of Jilin University,Changchun ,China.Study of Solid-Phase Time-Resolved Fluorescence Label Immunoassay[J].Spectroscopy and Spectral Analysis,2004,24(12):1601-1604. |
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| Authors: | PAN Li-hua ZHOU Shi-hong SUN Wen-wei XIE Wen-bing ZHAO Chao National Analytical Research Center of Electrochemistry and Spectroscopy Key Laboratory of Rare Earth Chemistry and Physics Changchun Institute of Applied Chemistry Chinese Academy of Sciences Changchun China China-Japan Union Hospital of Jilin University Changchun China |
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| Institution: | National Analytical Research Center of Electrochemistry and Spectroscopy, Key Laboratory of Rare Earth Chemistry and Physics, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, China. |
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| 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. |
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| Keywords: | Solid-phase time-resolved fluorimmunoassay Europium labels Anti-hepatits B surface 4 7-bis-chorosulfophenyl-1 10-phenanthroline-2 9-dicarboxylic acid |
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