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A. V. Petrakova, A. E. Urusov, A. V. Zherdev, B. B. Dzantiev

Comparison of techniqies for antibodies immobilization on the surface of magnetic particles in pseudo-homogeneous immunoenzyme assay of aflatoxin B1

Abstract

Pseudo-homogeneous enzyme immunoassay (EIA) of aflatoxin B1 (AFB1) has been realized with the use of conjugates between monoclonal antibodies against AFB1and magnetic particles (MP). The assay includes concentration of AFB1 from the tested sample on the surface of the MP-antibody conjugate, binding of AFB1-peroxidase conjugate with free sites of the antibodies, separation of formed complexes from unreacted components by magnetic field, and registration of enzymatic activity for peroxidase label bound with the MP. The preparations of antibody conjugates were obtained by three ways, physical adsorption on native MP and covalent binding to MP covers formed by oleic acid or polystyrene, and compared. EIA for these conjugates has limits of AFB1 detection equal to 2.6; 0.4 and 0.6 ng/mL, respectively. The pseudo-homogeneous EIA format enables reduction of immunoreagents incubation to 5 min (total duration of the analysis is 20 min) and allows to concentrate the analyte from the tested samples. These features determine advantages of the proposed assay as a tool for high sensitive control of toxic contaminants in food stuffs.
Key words: magnetic nanoparticles, immunoenzyme assay, immobilization of antibodies, aflatoxin B1.
Moscow University Chemistry Bulletin.
2015, Vol. 56, No. 6, P. 372
   

Copyright (C) Chemistry Dept., Moscow State University, 2002
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