Chemiluminescent enzyme immunoassay for alpha-fetoprotein using beta-D-galactosidase as label and 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside as substrate.
We developed a sensitive chemiluminescent sandwich-type enzyme immunoassay (CL-EIA) of alpha-fetoprotein (AFP) using beta-D-galactosidase (beta-gal) as a label and 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside as a substrate. The CL-EIA for AFP was performed using two monoclonal antibodies, one antibody is labelled with beta-gal, the other is coated onto the inside surface of a polystyrene tube. The detection limit for AFP was 0.5 ng/mL, equivalent to 10 pg/assay tube. The coefficient of variation for within and between assay imprecision were 2.0%-4.9% (n = 10) and 4.4%-9.8% (n = 5), respectively. AFP values in serum determined by this method correlated well with those obtained by radioimmunoassay (n = 26, r = 0.99). This sensitive AFP assay can be performed within 4 h and can be used as a routine assay in clinical diagnosis.
Arakawa
H
H
School of Pharmaceutical Sciences, Showa University, Tokyo, Japan.
Ikegami
T
T
Maeda
M
M
Tsuji
A
A
D006528:Carcinoma, Hepatocellular / Q000097:blood; D005697:Galactosides; D006505:Hepatitis / Q000097:blood; D006801:Humans; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G; D007202:Indicators and Reagents; D007211:Indoles; D008103:Liver Cirrhosis / Q000097:blood; D008107:Liver Diseases / Q000097:blood*; D008113:Liver Neoplasms / Q000097:blood; D008163:Luminescent Measurements; D008832:Microchemistry; D011863:Radioimmunoassay / Q000379:methods; D012016:Reference Values; D000509:alpha-Fetoproteins / Q000032:analysis*; D001616:beta-Galactosidase
D005697:Galactosides; D007074:Immunoglobulin G; D007202:Indicators and Reagents; D007211:Indoles; D000509:alpha-Fetoproteins; D001616:beta-Galactosidase; C044888:5-bromo-4-chloro-3-indolyl beta-galactoside