Comparative study of strategies for antibody immobilization onto the surface of magnetic particles in pseudo-homogeneous enzyme immunoassay of aflatoxin B1

被引：1
作者：
Petrakova A.V. [1 ]
Urusov A.E. [1 ]
Zherdev A.V. [1 ]
Dzantiev B.B. [1 ]
机构：
[1] Bach Institute of Biochemistry, Research Centre of Biotechnology, Russian Academy of Sciences, Moscow
来源：
Moscow University Chemistry Bulletin | 2016年 / 71卷 / 1期
关键词：
aflatoxin B1; antibody immobilization; enzyme immunoassay; magnetic particles;
D O I：
10.3103/S0027131416010119
中图分类号：
学科分类号：
摘要：
Pseudo-homogeneous enzyme immunoassay (EIA) of aflatoxin B1 (AFB1) was accomplished using anti-AFM1 monoclonal antibodies conjugated with magnetic particles (MPs). The assay includes the concentration of AFB1 from the test sample on the surface of the MP-antibody conjugate, the binding of the AFB1-peroxidase conjugate to free sites of the antibodies, the separation of the complexes that formed from unreacted components by means of magnetic field, and the evaluation of the enzymatic activity of MP-bound peroxidase. A comparative study of antibody conjugates, which were prepared by three different methods, namely, by physical adsorption on native MP and covalent binding to oleic acidor polystyrene-coated MPs, was performed. For these conjugates, the detection limits of EIA for AFB1 are 2.6, 0.4, and 0.6 ng/mL, respectively. The advantages of the pseudo-homogeneous EIA format as a tool for the highly sensitive control of toxic contaminants in food are the shorter time of incubation of immunoassay reagents (5 min; the total assay time is 20 min) and the possibility of concentrating the analyte from the test samples. © 2016, Allerton Press, Inc.
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页码：48 / 53
页数：5
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