[Study of solid-phase time-resolved fluorescence label immunoassay].
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.
Pan
Li-hua
LH
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.
Zhou
Si-hong
SH
Sun
Wen-wei
WW
Xie
Wen-bing
WB
Zhao
Chao
C
D005453:Fluorescence; D005456:Fluorescent Dyes; D015200:Fluoroimmunoassay / Q000379:methods*; D010618:Phenanthrolines / Q000737:chemistry; D013050:Spectrometry, Fluorescence / Q000379:methods*
C519844:1,10-phenanthroline-2,9-dicarboxylic acid; D005456:Fluorescent Dyes; D010618:Phenanthrolines; C025205:1,10-phenanthroline