Evaluation of alpha-fetoprotein (AFP) in human serum by chemiluminescence enzyme immunoassay with magnetic particles and coated tubes as solid phases.
In this work, the monoclonal antibodies (McAbs) to alpha-fetoprotein (AFP) were immobilized on two different solid phases, i.e., magnetic particles (MP) and coated tubes (CT). Based on this, a MP based chemiluminescence enzyme immunoassay (MP-CLEIA) and a CT based CLEIA (CT-CLEIA) were proposed for the evaluation of AFP in human serum and their analytical merits were studied and compared. By detailed discussion of several performance variants, including the concentration of immobilized McAb, dilution ratio of horseradish peroxidase (HRP) labeled McAb (HRP-McAb), total assay time, substrate volume, chemiluminescent kinetics, and hook effect concentration, the advantages of MP-CLEIA became conspicuously apparent. Moreover, in the presence of MP, the catalytic activity of labeled enzyme was kept to high extent and the stability of immunoreagents was satisfied. Finally, 59 human serum samples were detected by the MP-CLEIA and a good correlation was obtained when comparing the results with that from a commercial electrochemiluminescence immunoassay kit.
Zhang
Qianyun
Q
School of Science, Beijing University of Chemical Technology, Beijing 100029, China.
Wang
Xu
X
Li
Zhenjia
Z
Lin
Jin-Ming
JM
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism; D001774:Blood Chemical Analysis / Q000295:instrumentation* / Q000379:methods*; D002417:Cattle; D003429:Cross Reactions; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism; D006735:Horseradish Peroxidase / Q000378:metabolism; D006801:Humans; D007124:Immunoenzyme Techniques / Q000295:instrumentation* / Q000379:methods*; D007700:Kinetics; D008163:Luminescent Measurements; D008280:Magnetics; D012680:Sensitivity and Specificity; D013194:Staining and Labeling; D013997:Time Factors; D000509:alpha-Fetoproteins / Q000032:analysis* / Q000276:immunology
D000911:Antibodies, Monoclonal; D000509:alpha-Fetoproteins; D006735:Horseradish Peroxidase; D016650:Fluorescein-5-isothiocyanate