2-9-dicarboxy-1-10-phenanthroline and 1-10-phenanthroline

The authors labeled bovine serum albumin (BSA) with a new europium chelator 4,7-bis(chlorosulfophenyl)-1,10-phenanthroline-2,9-dicarboxylic acid (BCPDA) which was synthesized by solid phase time-resolved fluorescence immunoassay (TRFIA) technology. The process of BCPDA-labeling-BSA reaction was studied. Using Coomassie brilliant blue method, concentrations of BSA and BCPDA were determined in BCPDA-BSA labeled sample. The labeling reaction condition was studied. Labeling ratio and protein labeling recovery were calculated. BCPDA-BSA-Eu3+ complex has a very large Stokes shift (270 nm) and can emit very strong fluorescence band at 611.2 nm which is very narrow (10 nm). BCPDA-BSA-Eu3+ complex exhibits very long fluorescence lifetimes by the experiment demonstration. Also, BCPDA and protein-BCPDA-Eu3+ complex is relatively stable.

Topics: Animals; Cattle; Europium; Fluoroimmunoassay; Phenanthrolines; Proteins; Serum Albumin, Bovine; Spectrometry, Fluorescence

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.

Topics: Fluorescence; Fluorescent Dyes; Fluoroimmunoassay; Phenanthrolines; Spectrometry, Fluorescence