Study of Solid-Phase Time-Resolved Fluorescence Label Immunoassay
PAN Li-hua1, ZHOU Shi-hong1，SUN Wen-wei2，XIE Wen-bing1，ZHAO Chao1
1. 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 2. China-japan Union Hospital of Jilin University, Changchun 130031, China
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-Eu3+ complex is stable. It can emit very high fluorescence intensity with very long fluorescence life times. The fluorescence of Protein-BCPDA-Eu3+ 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
[1] Soini E, Hemmila I. Clin. Chem.，1979, 25: 353. [2] Hemmila I. Clin. Chem.，1985, 31：359. [3] Evangelista R A, Pollak A, Allore B et al. Clin. Biochem., 1988，21：173. [4] Mathis G. J. Clin. Ligand Assay．，1997, 20：141. [5] Yuan Jingli，Matsumoto Kazuko, Kimura Hiroko. Anal. Chem．，1998, 70：596. [6] PAN Li-hua，GAO Min, LIN Min, DING Chun-li, SUN Wen-wei, ZHAO Jie(潘利华，高 敏，林 敏，丁春黎，孙文伟，赵 劼). Spectroscopy and Spectral Analysis(光谱学与光谱分析)，2002，22(4)：573. [7] PAN Li-hua, ZENG Chun-hui, GUAN Ming, ZHAO Chao, LIN Min, XIE Wen-bing(潘利华，曾春慧，关 铭，赵 超，林 敏，谢文兵). Spectroscopy and Spectral Analysis(光谱学与光谱分析), 2003，23(5)：903.
