Fluorescence immunoassay of alpha-fetoprotein with iron(III) tetrasulfonatophthalocyanine as a mimetic enzyme labeling reagent.
A new fluorimetric immunoassay for alpha-fetoprotein (AFP) has been developed using a novel promising mimetic peroxidase, iron(III) tetrasulfonatophthalocyanine (FeTSPc), as a labeling reagent to catalyze the fluorescence reaction of P- hydroxyphenylacetic acid (P-HPA) and hydrogen peroxide (H2O2). In the competitive immunoassay, anti-AFP antibody was coated on a 96-well plate (polystyrene) and a constant amount of FeTSPc-labeled AFP and a known amount of test solution were added. Non-labeled and FeTSPc-labeled AFP compete for binding to the plate-bound antibody. After the immunoreaction, the immunochemically adsorbed FeTSPc-AFP conjugate moiety was determined by measuring the fluorescence produced in a solution containing P-HPA and H2O2. AFP can be determined in the concentration range of 1-300 ng mL(-1) with a detection limit of 0.5 ng mL(-1).
Yang
H H
HH
Department of Chemistry, Xiamen University, P.R. China.
Zhu
Q Z
QZ
Li
D H
DH
Lin
P
P
Ding
M T
MT
Xu
J G
JG
D000906:Antibodies; D000936:Antigen-Antibody Complex; D001667:Binding, Competitive; D004306:Dose-Response Relationship, Immunologic; D005290:Ferric Compounds / Q000138:chemical synthesis* / Q000737:chemistry; D015200:Fluoroimmunoassay / Q000379:methods*; D006735:Horseradish Peroxidase; D006801:Humans; D008665:Metalloporphyrins / Q000138:chemical synthesis* / Q000737:chemistry; D010648:Phenylacetates / Q000737:chemistry; D000509:alpha-Fetoproteins / Q000032:analysis* / Q000276:immunology
D000906:Antibodies; D000936:Antigen-Antibody Complex; D005290:Ferric Compounds; D008665:Metalloporphyrins; D010648:Phenylacetates; D000509:alpha-Fetoproteins; C430149:iron(III) tetrasulfonatophthalocyanine; C008070:4-hydroxyphenylacetic acid; D006735:Horseradish Peroxidase