Enhancement of the sensitivity of a capillary electrophoresis immunoassay for estradiol with laser-induced fluorescence based on a fluorescein-labeled secondary antibody.
A competitive immunoassay for estradiol (E2) based on capillary electrophoresis was established. This method was based on the competitive reaction of complete antigen and E2 with a limited amount of monoclonal antibody, with fluorescein isothiocyanate (FITC)-labeled secondary antibody as a fluorescence probe. The addition of the thermally reversible hydrogel, poly-N-isopropylacrylamide (pNIPA) in the buffer as a replaceable packing material improved reproducibility and resolution of the method. This capillary electrophoresis immunoassay with laser-induced fluorescence can be applied to determine E2 with good precision at concentrations as low as 9 pg/mL. Details of typical separations of immunological complex and free reactants are presented.
Wang
Y C
YC
Department of Chemical Biology, College of Chemistry, Peking University, Beijing, China.
Su
P
P
Zhang
X X
XX
Chang
W B
WB
D000911:Antibodies, Monoclonal; D019075:Electrophoresis, Capillary / Q000379:methods / Q000592:standards; D004958:Estradiol / Q000032:analysis* / Q000276:immunology; D019793:Fluorescein; D005456:Fluorescent Dyes; D006801:Humans; D007118:Immunoassay / Q000379:methods / Q000592:standards; D007834:Lasers; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D005456:Fluorescent Dyes; D004958:Estradiol; D019793:Fluorescein