Chemiluminescent enzyme immunoassay using beta-D-galactosidase as the label and the bis(2,4,6-trichlorophenyl)oxalate-fluorescent dye system.
A chemiluminescent enzyme immunoassay (EIA) using beta-D-galactosidase as the enzyme label is described in this report. The activity of beta-D-galactosidase, after separation of bound and free fractions, was measured using a coupled enzyme procedure: lactose and glucose oxidase were used as the substrate and coupling enzyme, respectively. Hydrogen peroxide generated from glucose was determined by the chemiluminescence reaction using the bis(2,4,6-trichlorophenyl)oxalate-fluorescent dye system. This assay system was applied to the EIA of phenytoin using beta-D-galactosidase as the label and assay sensitivity was increased about 10 times compared with the original method. This method may also be applied to other EIAs of digitoxin, alpha-fetoprotein and thyroid-stimulating hormone.
S
S
Takayasu
M
M
Maeda
A
A
Tsuji
D004074:Digitoxin / Q000032:analysis; D006863:Hydrogen-Ion Concentration; D007124:Immunoenzyme Techniques; D007700:Kinetics; D008163:Luminescent Measurements; D009994:Osmolar Concentration; D010070:Oxalates; D010672:Phenytoin / Q000032:analysis; D013696:Temperature; D013972:Thyrotropin / Q000032:analysis; D000509:alpha-Fetoproteins / Q000032:analysis; D001616:beta-Galactosidase
D010070:Oxalates; D000509:alpha-Fetoproteins; C036521:bis(2,4,6-trichlorophenyl)oxalate; D010672:Phenytoin; D013972:Thyrotropin; D004074:Digitoxin; D001616:beta-Galactosidase