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1

Bach Institute of Biochemistry of the Russian Academy of Sciences, Leninsky Prospect 33, Moscow 119071, Russia

2

IL Test-Pushchino Ltd., Gruzovaya Street 1g, Pushchino 142290, Moscow Region, Russia



These authors contributed equally to this work.





*

Author to whom correspondence should be addressed.



Abstract The main limitations of microplate-based enzyme immunoassays are the prolonged incubations necessary to facilitate heterogeneous interactions, the complex matrix and poorly soluble antigens, and the significant sample dilutions often required because of the presence of organic extractants. This study presents the use of antibody immobilization on the surface of magnetic particles to overcome these limitations in the detection of the mycotoxin, aflatoxin B1. Features of the proposed system are a high degree of nanoparticle dispersion and methodologically simple immobilization of the antibodies by adsorption. Reactions between the immobilized antibodies with native and labeled antigens are conducted in solution, thereby reducing the interaction period to 5 min without impairing the analytical outcome. Adsorption of immunoglobulins on the surface of magnetic nanoparticles increases their stability in aqueous-organic media, thus minimizing the degree of sample dilution required. Testing barley and maize extracts demonstrated a limit of aflatoxin B1 detection equal to 20 pg-mL and total assay duration of 20 min. Using this method, only the 3-fold dilution of the initial methanol-water 60-40 extraction mixture in the microplate wells is necessary. The proposed pseudo-homogeneous approach could be applied toward immunodetection of a wide range of compounds. View Full-Text

Keywords: immunoassay; magnetic particles; mycotoxins; aflatoxin immunoassay; magnetic particles; mycotoxins; aflatoxin





Autor: Alexandr E. Urusov 1,†, Alina V. Petrakova 1,†, Maxim V. Vozniak 2, Anatoly V. Zherdev 1 and Boris B. Dzantiev 1,*

Fuente: http://mdpi.com/



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