On the basis of Eu(III)-4,7-bis(chlorosulfophenyl)-1,10-phenanthroline-2,9-di-carboxylic
acid chelate (
BCPDA) that was synthesized and characterized for time-resolved fluoroimmunoassay (TRFIA), Donkey anti-
hepatitis B surface (anti-HBs) was labeled with
BCPDA-Eu3+.
Coomassie Brilliant Blue was used to determine the
protein concentration and radio immunoassay (RIA) for detecting the
biological activity in the labeled
protein. Optimal conditions for the
protein labeling were obtained by monitoring the reaction. Results suggested that the
protein could be labeled with
BCPDA under relatively moderate conditions. As a practical application, a
protein-
BCPDA-Eu3+ chelate was obtained by using
BCPDA-
protein that reacted with EuCl3 under certain conditions. Some properties of
BCPDA and
protein-
BCPDA-Eu3+, such as absorption spectrum, emission spectrum and fluorescence lifetime, were discussed. The detection limit and the linear working range of the established method were also investigated.