Determination of alpha-fetoprotein concentration based on liquid-phase binding assay using anion exchange chromatography and sulfated peptide introduced antibody.
A new immunoassay for alpha-fetoprotein based upon liquid-phase binding reactions is described. In this procedure, a sample that contains alpha-fetoprotein is mixed with a solution of two anti-alpha-fetoprotein monoclonal antibodies complexed with peroxidase and sulfated peptide, respectively. After incubation, immune complex is separated from other components by anion exchange column chromatography. Immune complex is quantified using fluorometric detection by peroxidase enzymatic activity. Peroxidase activity correlated with a alpha-fetoprotein with a 1:1 relationship. The stoichiometric immunoreaction allowed a large analytical range (0.4-7500 ng/ml) with a linear dose-response relationship, high sensitivity and good precision. Endogenous substances did not interfere with assay performance. Assay results showed good correlation with other established methods. These results indicate that the method is useful for clinical alpha-fetoprotein determinations.
Osaka Research Laboratories, Wako Pure Chemical Industries, Amagasaki, Hyogo, Japan.
Y
Y
Yamagata
H
H
Katoh
K
K
Nakamura
T
T
Tanaka
S
S
Satomura
S
S
Matsuura
D000838:Anions; D002852:Chromatography, Ion Exchange / Q000379:methods*; D005470:Fluorometry; D007118:Immunoassay / Q000379:methods*; D007124:Immunoenzyme Techniques; D011485:Protein Binding; D012680:Sensitivity and Specificity; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000276:immunology; D000509:alpha-Fetoproteins / Q000032:analysis* / Q000276:immunology
D000838:Anions; D000509:alpha-Fetoproteins; C005662:tyrosine O-sulfate; D014443:Tyrosine